To see the other types of publications on this topic, follow the link: In vitro cultured.
Journal articles on the topic 'In vitro cultured'
Create a spot-on reference in APA, MLA, Chicago, Harvard, and other styles
Consult the top 50 journal articles for your research on the topic 'In vitro cultured.'
Next to every source in the list of references, there is an 'Add to bibliography' button. Press on it, and we will generate automatically the bibliographic reference to the chosen work in the citation style you need: APA, MLA, Harvard, Chicago, Vancouver, etc.
You can also download the full text of the academic publication as pdf and read online its abstract whenever available in the metadata.
Browse journal articles on a wide variety of disciplines and organise your bibliography correctly.
1
Nishii, R., K. Imai, H. Koyama, and O. Dochi. "166 EFFECT OF INDIVIDUAL CULTURE SYSTEM ON IN VITRO DEVELOPMENT OF IN VITRO-MATURED - IN VITRO-FERTILIZED BOVINE EMBRYOS." Reproduction, Fertility and Development 24, no. 1 (2012): 195. http://dx.doi.org/10.1071/rdv24n1ab166.
Full textAbstract:
An individual in vitro culture system for bovine embryo needs to be developed for the study of embryo developmental competence. The objective of this study was to examine the effect of individual culture systems on the development of in vitro-matured–in vitro-fertilized bovine embryos. Two individual culture systems were compared. Cumulus–oocyte complexes (COC) were collected by aspiration of ovarian follicles (diameter, 2 to 5 mm) obtained from a local abattoir. The COC were matured in TCM-199 supplemented with 5% calf serum and 0.02 AU mL–1 of FSH. Groups of 20 COC were incubated in 100-μL droplets of IVM media at 38.5°C under an atmosphere of 5% CO2 for 20 h. After 18 h of gamete co-culture (3 × 106 sperm mL–1), the presumptive zygotes were cultured in CR1aa medium supplemented with 5% calf serum at 38.5°C under an atmosphere of 5% CO2, 5% O2 and 90% N2 for 9 days (fertilization = Day 0). The presumptive zygotes were randomly assigned to 1 of the following 3 treatments: single culture (1 zygote was cultured in a 5-μL droplet), well-of-the-well (WOW; Sugimura et al. 2010 Biol. Reprod. 83, 970–978) culture (25 zygotes were cultured individually in each 125-μL droplet) and control culture (25 zygotes were cultured in a 125-μL droplet). Embryo development was evaluated for cleavage and blastocyst rates, on Day 2 and Day 7 to 9 after IVF, respectively. The rates of development up to the blastocyst stage and total cell number in the blastocysts, determined by an air-drying method, were investigated. The cleavage and blastocyst rates were analysed by the chi-square test and the total cell numbers were analysed by ANOVA. The cleavage rates were significantly higher in the control and WOW groups than in the single-culture group (P < 0.01) and the blastocyst rates were significantly lower in the single-culture group than in the control culture group (P < 0.05; Table 1). The total cell numbers (mean ± s.d.) of blastocysts did not significantly differ between the single culture (154.6 ± 21.8), control culture (155.2 ± 22.5) and WOW culture (159.8 ± 27.0) groups. These results indicate that although the blastocyst rate was lower in the single-culture system than in the WOW or group culture system, in vitro-matured–in vitro-fertilized bovine embryos can be cultured using the single-culture system. Moreover, the quality of blastocysts developed by the single-culture system is the same as that of blastocysts developed using the other 2 culture systems. Table 1.Effect of different culture methods for bovine embryo development
2
du Preez, Francois, Antoinette Paula Malan, and Pia Addison. "Potential of in vivo- and in vitro-cultured entomopathogenic nematodes to infect Lobesia vanillana (Lepidoptera: Tortricidae) under laboratory conditions." PLOS ONE 16, no. 8 (2021): e0242645. http://dx.doi.org/10.1371/journal.pone.0242645.
Full textAbstract:
Entomopathogenic nematodes (EPNs) have been successfully applied as biological control agents against above ground and soil stages of insect pests. However, for commercial application, it is crucial to mass culture these nematodes using in vitro liquid culture technology, as it is not attainable when using susceptible insects as hosts. Lobesia vanillana (Lepidoptera: Tortricidae) is regarded a sporadic pest of wine grapes in South Africa. The in vivo- and in vitro-cultured South African EPNs, Steinernema yirgalemense and Steinernema jeffreyense (Rhabditida: Steinernematidae), were evaluated against larvae and pupae of L. vanillana in laboratory bioassays. For larvae, high mortality was observed for all treatments: In vitro-cultured S. yirgalemense (98%) performed better than S. jeffreyense (73%), while within in vivo cultures, there was no difference between nematode species (both 83%). No significant difference was detected between in vivo- and in vitro cultures of the same nematode species. The LD50 of the in vitro-cultured S. yirgalemense, was 7.33 nematodes per larva. Mortality by infection was established by dissecting L. vanillana cadavers and confirming the presence of nematodes, which was > 90% for all treatments. Within in vitro cultures, both S. yirgalemense and S. jeffreyense were able to produce a new cohort of infective juveniles from L. vanillana larvae. Pupae, however, were found to be considerably less susceptible to EPN infection. This is the first study on the use of EPNs to control L. vanillana. The relative success of in vitro-cultured EPN species in laboratory assays, without any loss in pathogenicity, is encouraging for further research and development of this technology.
3
Jarrahy, Reza, Weibiao Huang, George H. Rudkin, et al. "Osteogenic differentiation is inhibited and angiogenic expression is enhanced in MC3T3-E1 cells cultured on three-dimensional scaffolds." American Journal of Physiology-Cell Physiology 289, no. 2 (2005): C408—C414. http://dx.doi.org/10.1152/ajpcell.00196.2004.
Full textAbstract:
Osteogenic differentiation of osteoprogenitor cells in three-dimensional (3D) in vitro culture remains poorly understood. Using quantitative real-time RT-PCR techniques, we examined mRNA expression of alkaline phosphatase, osteocalcin, and vascular endothelial growth factor (VEGF) in murine preosteoblastic MC3T3-E1 cells cultured for 48 h and 14 days on conventional two-dimensional (2D) poly(l-lactide-co-glycolide) (PLGA) films and 3D PLGA scaffolds. Differences in VEGF secretion and function between 2D and 3D culture systems were examined using Western blots and an in vitro Matrigel-based angiogenesis assay. Expression of both alkaline phosphatase and osteocalcin in cells cultured on 3D scaffolds was significantly downregulated relative to 2D controls in 48 h and 14 day cultures. In contrast, elevated levels of VEGF expression in 3D culture were noted at every time point in short- and long-term culture. VEGF protein secretion in 3D cultures was triple the amount of secretion observed in 2D controls. Conditioned medium from 3D cultures induced an enhanced level of angiogenic activity, as evidenced by increases in branch points observed in in vitro angiogenesis assays. These results collectively indicate that MC3T3-E1 cells commit to osteogenic differentiation at a slower rate when cultured on 3D PLGA scaffolds and that VEGF is preferentially expressed by these cells when they are cultured in three dimensions.
4
Silva, J. R. V., T. Tharasanit, M. A. M. Taverne, et al. "The activin-follistatin system and in vitro early follicle development in goats." Journal of Endocrinology 189, no. 1 (2006): 113–25. http://dx.doi.org/10.1677/joe.1.06487.
Full textAbstract:
The aim of the present study was to investigate the effects of activin-A and follistatin on in vitro primordial and primary follicle development in goats. To study primordial follicle development (experiment 1), pieces of ovarian cortex were cultured in vitro for 5 days in minimal essential medium (MEM) supplemented with activin-A (0, 10 or 100 ng/ml), follistatin (0, 10 or 100 ng/ml) or combinations of the two. After culture, the numbers of primordial follicles and more advanced follicle stages were calculated and compared with those in non-cultured tissue. Protein and mRNA expression of activin-A, follistatin, Kit ligand (KL), growth differentiation factor-9 (GDF-9) and bone morphogenetic protein-15 (BMP-15) in non-cultured and cultured follicles were studied by immunohistochemistry and PCR. To evaluate primary follicle growth (experiment 2), freshly isolated follicles were cultured for 6 days in MEM plus 100 ng/ml activin-A, 100 ng/ml follistatin or 100 ng/ml activin-A plus 200 ng/ml follistatin. Morphology, follicle and oocyte diameters in cultured tissue and isolated follicles before and after culture were assessed. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) reactions were performed to study DNA fragmentation in follicles. In experiment 1, it was found that goat primordial follicles were activated to develop into more advanced stages, i.e. intermediate and primary follicles, during in vitro culture, but neither activin-A nor follistatin affected the number of primordial follicles that entered the growth phase. Activin-A treatment enhanced the number of morphologically normal follicles and stimulated their growth during cortical tissue culture. The effects were, however, not counteracted by follistatin. The follicles in cultured goat tissue maintained their expression of proteins and mRNA for activin-A, follistatin, KL, GDF-9 and BMP-15. Fewer than 30% of the atretic follicles in cultured cortical tissue had TUNEL-positive (oocyte or granulosa) cells. Activin-A did not affect the occurrence of TUNEL-positive cells in follicles within cortical tissue. In experiment 2, addition of activin-A to cultured isolated primary follicles significantly stimulated their growth, the effect being counteracted by follistatin. Absence of such a neutralizing effect of follistatin in the cultures with ovarian cortical tissue can be due to lower dose of follistatin used and incomplete blockage of activin in these experiments. In contrast to cortical enclosed atretic follicles, all atretic follicles that had arisen in cultures with isolated primary follicles had TUNEL-positive cells, which points to differences between isolated and ovarian tissue-enclosed follicles with regard to the followed pathways leading to their degeneration. In summary, this in vitro study has demonstrated that cultured goat primordial follicles are activated to grow and develop into intermediate and primary follicles. During in vitro culture, the follicles maintain their ability to express activin-A, follistatin, KL, GDF-9 and BMP-15. The in vitro growth and survival of activated follicles enclosed in cortical tissue and the in vitro growth of isolated primary follicles are stimulated by activin-A.
5
Kobayashi, Tsunehiro, Hossein Arefanian, George Harb, et al. "Prolonged Survival of Microencapsulated Neonatal Porcine Islet Xenografts in Immune-Competent Mice without Antirejection Therapy." Cell Transplantation 17, no. 10-11 (2008): 1243–56. http://dx.doi.org/10.3727/096368908787236602.
Full textAbstract:
Several studies have demonstrated that in vitro culture of islets prolonged islet graft survival in immune-competent mice without administration of antirejection drugs. However, we recently showed that in vitro cultured microencapsulated neonatal porcine islets (NPI) were rejected in immune-competent mice not receiving antirejection therapy. The aim of this study was to determine whether culture of microencapsulated NPI in vivo could promote long-term survival of microencapsulated NPI in immune-competent mice without administration of antirejection drugs. Microencapsulated NPI that were cultured in vitro for 7 and 50 days or transplanted initially in immune-deficient C.B.-17 SCID-BEIGE mice for 100 days (in vivo cultured) were characterized and transplanted into streptozotocin-induced diabetic immune-competent BALB/c mice. Day 50 in vitro cultured and day 100 in vivo cultured microencapsulated NPI showed significantly higher insulin and DNA content, indicating maturation of NPI compared to day 7 in vitro cultured microencapsulated NPI. Interestingly, in vivo cultured microencapsulated NPI expressed lower levels of porcine antigens compared to day 7 and day 50 in vitro cultured microencapsulated NPI. Transplantation of day 7 in vitro cultured microencapsulated NPI did not reverse diabetes in immune-competent BALB/c mouse recipients. In contrast, transplantation of day 50 in vitro cultured and in vivo cultured microencapsulated NPI into diabetic immune-competent BALB/c mice resulted in the immediate reversal of hyperglycemia within 2 days posttransplantation. However, all recipients of day 50 in vitro cultured microencapsulated NPI eventually rejected their grafts by day 15 posttransplantation, while 6 of 10 BALB/c mouse recipients of in vivo cultured microencapsulated NPI maintained normoglycemia for 100 days posttransplantation. These results show that in vivo culture of NPI in immune-deficient mice results in the modulation of NPI, which allows for their long-term survival in immune-competent mice without antirejection therapy.
6
Wilson, Timothy, Reeta Viitala, Mervi Puska, Mika Jokinen, and Risto Penttinen. "Macrophage Induced Effect of Particulate Silica on Rat Mesenchymal Stem Cells In Vitro." Key Engineering Materials 396-398 (October 2008): 123–26. http://dx.doi.org/10.4028/www.scientific.net/kem.396-398.123.
Full textAbstract:
The role of silica and macrophages in fibrosis is well documented, but in bone formation it is relatively unknown despite decades of research with bioactive glasses. In this study macrophages were isolated from rat peritoneal and then cultured for five days in the presence of two types of silica microparticles with different solubilities. After the fifth day the culture medium was collected, purified and used as an additive in bone marrow derived rat stem cell cultures. The stem cells were cultured for five days in α-mem containing only 0,5% of FCS, enabling cell survival but disrupting their proliferation. As controls, stem cells were also cultured in α-mem containing silica microparticles. At days one and five the amount of soluble collagen was assayed from the culture medium and the cells were counted. All stem cell cultures with macrophage medium additives were found to be proliferative, with statistically significant difference to controls. However, collagen was only produced in cultures containing medium from macrophages cultured with fast-dissolving silica microparticles. This suggests that silica can induce cell proliferation and extra cellular matrix protein secretion which is mediated by macrophages, and that the solubility of silica is also a major factor in this reaction.
7
Nas, Mehmet Nuri, Nedim Mutlu, and Paul E. Read. "Random Amplified Polymorphic DNA (RAPD) Analysis of Long-term Cultured Hybrid Hazelnut." HortScience 39, no. 5 (2004): 1079–82. http://dx.doi.org/10.21273/hortsci.39.5.1079.
Full textAbstract:
RAPD and phenotypic analysis were conducted to assess clonal stability of hazelnuts generated from axillary buds cultured in vitro for long-term. The nuts produced on in vitro-propagated plants were indistinguishable from those of donor plants. With the exception of rare horizontal (plagiotropic) growth, all in vitro-propagated plants exhibited phenotypes similar to those of donor plants. RAPD analysis did not reveal any somaclonal variation between donor plants from which in vitro cultures were initiated and micropropagated plants (6-year cultures), and no somaclonal variation was detected among in vitro-propagated plants. However, polymorphism (15.6%) was detected between the parent plant and its in vitro-propagated progenies (from seedlings). These results show a good discriminatory power of RAPD to detect polymorphism between samples where it is expected, and it can be effectively used for genetic assessment of micropropagated hazelnut. No evidence of genetic or epigenetic changes was observed in long-term cultured hazelnut, and thus long-term in vitro culture of hazelnut does not seem to limit its clonal propagation.
8
Racusen, L. C., B. A. Fivush, H. Andersson, and W. A. Gahl. "Culture of renal tubular cells from the urine of patients with nephropathic cystinosis." Journal of the American Society of Nephrology 1, no. 8 (1991): 1028–33. http://dx.doi.org/10.1681/asn.v181028.
Full textAbstract:
Nephropathic cystinosis represents a prototype for lysosomal storage diseases and is the most common cause of renal tubular Fanconi's syndrome. Mechanisms of the tubular transport defects in this disease have not been defined, however, in part because the cells readily cultured from affected patients, leukocytes and fibroblasts, do not express epithelial transport functions. Except for a single autopsy report, renal tubular cells from these patients have not been studied in vitro. In these studies, noninvasive harvesting and culture of renal tubular cells from the urine of patients with cystinosis is described. Cultures of renal tubular cells could be established from over 50% of the isolates which contained viable cells and which remained uncontaminated in vitro. Cells had an epithelial morphology in culture, and the majority of cultured cells expressed proximal tubular brush border marker enzyme. Cultured cells also expressed the storage defect in vitro, containing cystine levels up to 100 times those of normal cells. Cultured cells could be depleted of cystine by using the thiol cysteamine. This in vitro model system should be very useful for studying the mechanisms of renal tubular transport defects in this disease.
9
Al-Juboory, Karim H., and Jabar H. Al-Niami. "Adventitious Shoot and Plantlet Formation from Cultured Apple Leaf Explants." HortScience 31, no. 4 (1996): 629f—629. http://dx.doi.org/10.21273/hortsci.31.4.629f.
Full textAbstract:
Leaves of wild apple (Malus domestica Borkh) were excised from in vitro grown shoots transversely cut into halves and plated onto petri dishes containing regeneration media. Cultures were kept in the dark for three weeks before adventitious shoots were observed. Callus from leaf explants produced adventitious shoots after 3 months of in vitro culture. Callus were cultured on Nitsch and Nitsch medium supplemented with a range of BA (0.0–2.0 μm) and NAA (0.0–10 μm). BA at 10 μm combined with NAA (0.5 μm) proved most effective for stimulating shoot proliferation of cultured apple. Plantlets from tissue culture were easily transferred to the greenhouse environment.
10
Romberger, D. J., P. Pladsen, L. Claassen, M. Yoshida, J. D. Beckmann, and S. I. Rennard. "Insulin modulation of bronchial epithelial cell fibronectin in vitro." American Journal of Physiology-Lung Cellular and Molecular Physiology 268, no. 2 (1995): L230—L238. http://dx.doi.org/10.1152/ajplung.1995.268.2.l230.
Full textAbstract:
Fibronectin (Fn) is involved in the migration of epithelial cells in re-epithelialization of wounds. Epithelial cell-derived Fn is particularly potent as a chemotactic factor for bronchial epithelial cells (BECs) in vitro. Thus modulation of airway epithelial cell Fn may be a key aspect of airway repair. Insulin is both an important growth factor and known chemotactic factor for cultured BECs. We postulated that insulin may modulate Fn production of cultured BECs. We examined this hypothesis utilizing bovine BECs in culture with serum-free media with and without insulin. BECs grown in media without insulin released more Fn into culture supernatants and contained more Fn in cell layers than cells grown with insulin. Labeling of cells with [35S]methionine demonstrated an increase in new protein production and Fn mRNA expression was increased. Increased Fn in BEC cultures without insulin was associated with an increase in active transforming growth factor-beta (TGF-beta) release as measured by a standard bioassay. Increased BEC Fn in cultures without insulin was partially inhibited by exposure of cultures to TGF-beta antibody. Thus insulin appears to modulate BEC Fn production in vitro in part through a TGF-beta-dependent mechanism. Insulin may be involved in airway repair mechanisms through modulation of epithelial cell Fn production.
11
Guo, W., K. Kamiya, J. Cheng, and J. Toyama. "Changes in action potentials and ion currents in long-term cultured neonatal rat ventricular cells." American Journal of Physiology-Cell Physiology 271, no. 1 (1996): C93—C102. http://dx.doi.org/10.1152/ajpcell.1996.271.1.c93.
Full textAbstract:
A primary culture of neonatal ventricular myocytes isolated from day-old rats was established for investigating the changes in action potentials and ion currents over long periods. Cells at days 5 and 15 in culture were studied. These changes in vitro were compared with those in situ derived from the age-matched freshly isolated cells. During primary culture, quiescent cells demonstrated shortening of action potential durations (APD) resembling the developmental changes observed in situ. The beating cultured cells were not associated with APD shortening. Despite constant current amplitudes, the densities of Ca2+ currents (ICa) decreased in the quiescent cultures at later ages as a result of cell enlargement. ICa densities were maintained in the beating cultured and freshly isolated cells. Acceleration in the inactivation of ICa was observed during developments both in vitro and in situ. In addition, the densities of transient outward currents (Ito) tripled and doubled in the quiescent and beating cells during 15-day cultures. However, Ito in beating cultured cells made less contribution to APD in contrast to the quiescent cultured and freshly isolated myocytes. These findings demonstrate that electrophysiological properties differ between two types of long-term cultured cells. ICa densities remained constant in the beating cultures, suggesting that cell beating may be required for the maintenance of ICa density in developing cardiomyocytes.
12
Alexander, J. Steven, and Thomas M. Crisp. "Gonadotrophic regulation of prolactin mediated progesterone secretion in vitro." Acta Endocrinologica 110, no. 2 (1985): 251–56. http://dx.doi.org/10.1530/acta.0.1100251.
Full textAbstract:
Abstract. The effects of preincubating rat granulosa cells with FSH, LH, and Prl on subsequent Prl mediated progesterone secretion were investigated. Granulosa cells were isolated from ovarian follicles 50 h after injection of 5 IU PMSG and were then plated on poly-l-lysine coated coverslips in serum supplemented medium. Cells were preincubated for 24 h in the absence of hormones (control) or with the addition of either 0.25, 2.5, 25 ng/ml rat FSH or rat LH, or 1 μg/ml rat Prl. Following the preincubation period, cells were maintained for an additional 6 or 8 days in the presence or absence of 1 μg/ml Prl. When cells were preincubated with FSH or LH, only the two higher concentrations (2.5 and 25 ng/ml) stimulated significantly more progesterone secretion than control cultures during the 24 h preincubation period. For each series of preincubations, cells cultured for 6 or 8 days in the presence of Prl secreted significantly more progesterone at each day of culture than cells cultured without Prl. Cells preincubated and cultured with Prl secreted only 3–7-fold more progesterone than cells preincubated in control medium and then cultured with Prl. Preincubation with FSH or LH promoted a 20–45-fold increase in Prl mediated progesterone secretion compared to control preincubation cultures that also subsequently were cultured with Prl. The magnitude of Prl mediated progesterone secretion observed through 6 days of culturing was dose dependent on the preincubation concentration of FSH or LH. The establishment of an in vitro model system in which gonadotrophins enhance the responsiveness of granulosa cells to Prl in serum supplemented medium provides the opportunity for study of the regulatory mechanisms involved with the induction and maintenance of such responsiveness.
13
Shannon, J. M., S. D. Jennings, and L. D. Nielsen. "Modulation of alveolar type II cell differentiated function in vitro." American Journal of Physiology-Lung Cellular and Molecular Physiology 262, no. 4 (1992): L427—L436. http://dx.doi.org/10.1152/ajplung.1992.262.4.l427.
Full textAbstract:
We have investigated whether the loss of differentiated function observed in adult rat alveolar type II cells cultured on a substratum that promotes cell spreading and flattening represents a reversible phenotypic change. Cells were cultured for 4 and 8 days in association with fetal rat lung fibroblast feeder layers on either attached collagen gels, which promote the loss of differentiated function, or on floating collagen gels, which support differentiation. A fifth group of cultures were maintained as attached gels for 4 days, then detached and cultured as floating gels for the remaining 4 days. Expression of mRNAs for surfactant proteins A, B, and C, patterns of phospholipid biosynthesis, rates and patterns of protein synthesis, and cell morphology were evaluated as markers of differentiation. Without exception, detaching the gels after 4 days in culture resulted in significant recovery of differentiated characteristics, demonstrating that type II cells modulate differentiated function in response to the culture environment. The results are discussed in relation to the importance of normal cell architecture to normal cell function and to the possible in vitro progression of type II cells to type I cells.
14
da Silva, Cleidson Manoel Gomes, Simone Vieira Castro, Luciana Rocha Faustino, et al. "Activin-A promotes the development of goat isolated secondary follicles in vitro." Zygote 23, no. 1 (2013): 41–52. http://dx.doi.org/10.1017/s0967199413000294.
Full textAbstract:
SummaryThe role of activin-A in follicular development and on the mRNA expression levels of different genes in goat secondary follicles was evaluated. Goat secondary follicles (≥150 μm) were cultured for 18 days under control conditions or with the addition of either 50 or 100 ng/ml activin-A (Experiment 1). The mRNA levels for the genes that code for activin-A, ActR-IA, ActR-IB, ActR-IIA, ActR-IIB, follicle stimulating hormone receptor (FSH-R) and P450 aromatase were measured in each condition (Experiment 2). We observed that after 6 days of culture, the antrum formation rate was higher in cultures with added activin-A than in the cultured control (P < 0.05). The addition of 50 ng/ml activin-A increased the follicular growth rate in the final third of the culture (days 12–18), resulting in a higher percentage of meiosis resumption (P < 0.05). On day 6, the addition of activin-A (50 ng/ml) increased the levels of ActR-IA mRNA compared with the cultured control (P < 0.05). After 18 days, the addition of 50 ng/ml activin-A significantly increased the levels of its own mRNA compared with the non-cultured control. Moreover, this treatment reduced the mRNA levels of P450 aromatase in comparison with the cultured control (P < 0.05). Higher levels of P450 aromatase mRNA were found for both activin-A treatments compared with the non-cultured control (P < 0.05). No difference in estradiol levels was detected among any of the tested treatments. In conclusion, the addition of activin-A to culture medium stimulated early antrum formation as well as an increase in the daily follicular growth rate and the percentage of meiosis resumption.
15
Stimart, Dennis P., John C. Mather, and Kenneth R. Schroeder. "Shoot Proliferation and Rooting in Vitro of Pulmonaria." HortScience 33, no. 2 (1998): 339–41. http://dx.doi.org/10.21273/hortsci.33.2.339.
Full textAbstract:
Expanding shoot tips of Pulmonaria `Roy Davidson' and Pulmonaria saccharata `Margery Fish' were cultured in vitro on a modified Murashige and Skoog medium containing BA to establish proliferating cultures for use in comparing BA concentrations on shoot proliferation and rooting. The optimum level for shoot proliferation was 8.8 μm BA. Greatest rooting was on medium without BA. Genotype and time in culture influenced shoot and root counts. Chemical names used: N6-benzyladenine (BA)
16
Mrázová, Veronika, Tatiana Betáková, Marcela Kúdelová, et al. "Murine Gammaherpesvirus (MHV-68) Transforms Cultured Cells in vitro." Intervirology 58, no. 2 (2015): 69–72. http://dx.doi.org/10.1159/000370071.
Full textAbstract:
Human dermal fibroblasts and mouse NIH/3T3 cells acquired the transformed phenotype (‘criss-cross' pattern of growth) after infection with ultraviolet-irradiated murine gammaherpesvirus (MuHV-4 strain 68; MHV-68). These cells with changed phenotype could be serially cultured for 5-6 passages (35-40 days), and then they entered into crisis and most of them died. In a small number of cultures, however, foci of newly transformed cells appeared from which two stable cell lines were derived. After 6-9 cell culture passages of the MHV-68 transformed cell lines, MHV-68 DNA and virus antigen could be detected by PCR and immunofluorescence assay along with the disappearance of actin bundles, indicating that both transformed cell lines might be oncogenic.
17
Iida, Jin, Takafumi Yoshikawa, Kazuhide Miyazaki, Noriko Okumura, and Yoshinori Takakura. "Osteogenic Potential of Estriol-Treated Cultured Bone - In Vitro and In Vivo -." Key Engineering Materials 284-286 (April 2005): 651–54. http://dx.doi.org/10.4028/www.scientific.net/kem.284-286.651.
Full textAbstract:
Osseous tissue can be formed by culturing marrow cells with compounds such as dexamethasone and that a bone matrix cultured in this manner possesses BMP activity. We have reported that artificial bones with a high level of osteogenic potential can be prepared by culturing artificial bone materials with cultured osseous tissue. Here, in an attempt to develop activated cultured bone constructs with even greater osteogenic potential, the effects of the female hormone estriol on osteogenesis were investigated. Bone marrow cells were collected from the femur shafts of 7-week-old male Fischer rats, and subjected to primary and secondary cultures. During secondary culture with or without dexamethasone (Dx), 10-5, 10-6, 10-7, 10-8 or 10-9 M of estriol was added to a standard culture medium containing ascorbic acid and β-glycerophosphosphate. The alkaline phosphatase(ALP) activity and Ca levels were measured and statistically analyzed. There was a significant difference in ALP activity between the control group and the estriol groups, and ALP activity was the highest in the 10-7 and 10-8 M groups, being about 2.5 times higher than in the control group. Similar results were seen for Ca levels. Furthermore, in vivo study showed10-7M-estriol-treated-cultured bone/ceramic construct has significant high osteogenic potential when it is grafted into in vivo. Estriol has been reported to increase bone mass, and the results of the present study suggest that the osteogenic potential of cultured bone constructs can be more than doubled by adjusting the concentration of estriol in bone marrow cell culture. Therefore, the use of estriol may be able to facilitate osteogenesis in bone regeneration therapy.
18
Walum, Erik, Elisabeth Hansson, and Alan L. Harvey. "In Vitro Testing of Neurotoxicity." Alternatives to Laboratory Animals 18, no. 1_part_1 (1990): 153–79. http://dx.doi.org/10.1177/026119299001800118.1.
Full textAbstract:
Many of the toxic compounds that are at large in the environment represent a risk to our neuronal functions. Chemicals may have a direct or indirect effect on the nervous system and they may interfere with general biochemical properties or specific neuronal structures and processes. In this review, a brief presentation of the major neurotoxicological targets is given, together with a discussion of some aspects of the use of different in vitro models for screening purposes and mechanistic studies. It is believed that in vitro methods offer special opportunities for the development of new neurotoxicological assays, and that this development will mainly involve cultured model systems. Therefore, a presentation of nerve and glia tissue culture methods is given, followed by an overview of how information on the action of mercury and mercurials, excitotoxins and acrylamide has been obtained through the use of cultured cell models. It is concluded that the developmental potential in cell neurotoxicology lies within the areas of separation and identification of cells representative for different structures in the nervous system, co-cultivation of different cell types, in vivo/in vitro (ex vivo) procedures, chemically defined media, metabolic competent cultures of human cells and improved physiological conditions for cultivation and exposure.
19
Kancherla, S. Latha, and Prem L. Bhalla. "In Vitro Propagation of Pandoreas." HortScience 36, no. 2 (2001): 348–50. http://dx.doi.org/10.21273/hortsci.36.2.348.
Full textAbstract:
Pandoreas, Australian natives of horticultural significance, were successfully propagated using tissue culture. A protocol for rapid in vitro multiplication of commercial cultivars was developed using nodal segments cultured on Murashige and Skoog medium containing either BA or kinetin. Maximum shoot induction and number of shoots per explant for P. pandorana (Andrews) Steenis and P. jasminoides (Lindley) Schumann were on 8.8 μm BA and 4.6 μm kinetin. Higher levels of cytokinin in the medium inhibited shoot formation. Tissue-cultured shoots were rooted with IBA. This study demonstrates that Pandoreas can be successfully micropropagated. Chemical names used: 6-benzylaminopurine (BA); 3-indole butyric acid (IBA).
20
Ali-Ahmad, Masooma, and Harrison Hughes. "807 PB 141 LEAF EPICUTICULAR WAX OF IN VITRO GRAPE PLANTLETS TREATED WITH AND WITHOUT POLYETHYLENE GLYCOL." HortScience 29, no. 5 (1994): 548g—548. http://dx.doi.org/10.21273/hortsci.29.5.548g.
Full textAbstract:
Scanning electron microscopic (SEM) studies and gravimetric analysis of in vitro cultured leaf surfaces showed reduced epicuticular wax (EW) structurally and quantitatively as compared to greenhouse plants. However, leaves of in vitro plantlets subjected to polyethylene glycol-treatment (PEG) showed an increase in quantitative and structural EW which was similar to that of greenhouse plants. Furthermore, leaves initiated during in vitro culture and which persisted, when transferred to the greenhouse, showed an increase in structural wax as well as in amount, 30 days after transplanting in the greenhouse. Similarly, leaves newly-formed in the greenhouse from in vitro cultured plants developed more dense crystalline structure and greater levels of wax than those leaves observed immediately after removal from culture. A correlation between density of structural EW and amount of EW were observed in in vitro cultured, PEG-treated in vitro cultured and greenhouse grown leaves.
21
Andrade, Evelyn R., Robert van den Hurk, Lívia A. Lisboa, et al. "Effects of ascorbic acid on in vitro culture of bovine preantral follicles." Zygote 20, no. 4 (2012): 379–88. http://dx.doi.org/10.1017/s0967199412000056.
Full textAbstract:
SummaryThe objective of this study was to evaluate the effects of adding ascorbic acid to the media for in vitro culture of cattle ovarian fragments and to determine their effects on growth activation and viability of early-stage follicles. The ovarian cortex was divided into small fragments; one fragment was immediately fixed (control) and the other fragments were cultured in minimum essential medium (MEM) supplemented or not with various doses of ascorbic acid. Ovarian tissue was processed for histology, transmission electron microscopy (TEM) and immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA). Compared with control fragments, the percentage of primordial follicles was reduced (p < 0.05) and the percentage of growing follicles had increased (p < 0.05) in cultured cortical fragments, independent of the tested medium or incubation time. Furthermore, compared with control tissue, culture of ovarian cortex for 8 days reduced the percentages of healthy, viable follicles (p < 0.05), but not when cultures were supplemented with 25, 50 or 100 μg/ml of ascorbic acid. Ultrastructural and immunohistochemical analysis of 8 day cultured ovarian cortical fragments, however, showed the integrity and viability of follicles only when fragments were cultured in presence of 50 μg/ml of ascorbic acid. In conclusion, this study demonstrated that addition of ascorbic acid to MEM at a concentration of 50 μg/ml not only stimulates the activation of 8 day in vitro cultured cattle primordial follicles and subsequent growth of activated follicles, but also safeguards the viability of these early-stage follicles.
22
Lacasse, J., C. Kreft, J. Gutkowska, J. Genest, and M. Cantin. "Cultured juxtaglomerular cells: production and localization of renin." Canadian Journal of Physiology and Pharmacology 65, no. 7 (1987): 1409–15. http://dx.doi.org/10.1139/y87-221.
Full textAbstract:
Trypsin- or collagenase-dispersed renal cortical cells from newborn mice were filtered and cultured. The cultures comprised 25% juxtaglomerular cells as identified by immunocytochemistry. Renin was measured by radioimmunoassay in the culture medium and in the cells at various time intervals. This in vitro system was responsive to isoproterenol, which stimulated renin release in a dose-dependent manner.
23
Barroso, P. A. A., L. R. F. M. Paulino, B. R. Silva, et al. "Effects of dexamethasone on growth, viability and ultrastructure of bovine secondary follicles cultured in vitro." Zygote 28, no. 6 (2020): 504–10. http://dx.doi.org/10.1017/s0967199420000416.
Full textAbstract:
SummaryThis study aimed to evaluate the effects of dexamethasone on development, viability, antrum formation and ultrastructural integrity of bovine secondary follicles cultured in vitro for 18 days. Bovine ovaries were obtained from slaughterhouses and secondary follicles of ~150–200 µm diameter were isolated and cultured in the laboratory in TCM-199+ alone or supplemented with different concentrations of dexamethasone (1, 10, 100 and 1000 ng/ml). Follicle viability was evaluated after the culture period, using calcein-AM (viable) and ethidium homodimer (nonviable). Follicle diameters and antrum formation were evaluated at days 0, 6, 12 and 18. Before or after in vitro culture, follicles were fixed for histological and ultrastructural analysis. Follicle diameters were evaluated using analysis of variance and Kruskal–Wallis test, while chi-squared test was used to evaluate the percentage of viable follicles and antrum formation (P < 0.05). Follicles cultured for 6 days with all treatments increased their diameters significantly, but there was no significant difference between treatments at the end of the culture period. In vitro cultured follicles showed antral cavity formation at the end of the culture period, but no influence of dexamethasone was seen. Ultrastructural analysis showed that follicles cultured with dexamethasone (1, 10, 100 and 1000 ng/ml) had well preserved granulosa cells. However, oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone showed signs of degeneration. It can be concluded that follicles cultured in vitro in the presence of dexamethasone demonstrated continuous in vitro growth, but oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone had poor ultrastructure.
24
Orihuela, Carlos J., Rob Janssen, Christopher W. Robb, David A. Watson, and David W. Niesel. "Peritoneal Culture Alters Streptococcus pneumoniae Protein Profiles and Virulence Properties." Infection and Immunity 68, no. 10 (2000): 6082–86. http://dx.doi.org/10.1128/iai.68.10.6082-6086.2000.
Full textAbstract:
ABSTRACT We have examined the properties of Streptococcus pneumoniae cultured in the murine peritoneal cavity and compared its virulence-associated characteristics to those of cultures grown in vitro. Analysis of mRNA levels for specific virulence factors demonstrated a 2.8-fold increase in ply expression and a 2.2-fold increase in capA3 expression during murine peritoneal culture (MPC). Two-dimensional gels and immunoblots using convalescent-phase patient sera and murine sera revealed distinct differences in protein production in vivo (MPC). MPC-grown pneumococci adhered to A549 epithelial cell lines at levels 10-fold greater than those cultured in vitro.
25
Alves, A. M. C. V., R. N. Chaves, R. M. P. Rocha, et al. "Dynamic medium containing growth differentiation factor-9 and FSH maintains survival and promotes in vitro growth of caprine preantral follicles after long-term in vitro culture." Reproduction, Fertility and Development 25, no. 6 (2013): 955. http://dx.doi.org/10.1071/rd12180.
Full textAbstract:
The aim of the present study was to evaluate the effects of growth differentiation factor 9 (GDF-9) and FSH on the in vitro development of caprine preantral follicles cultured for 16 days. Ovarian fragments were cultured in αMEM+ (α-minimum essential medium, pH 7.2–7.4, 10 μg mL–1 insulin, 5.5 μg mL–1 transferrin, 5.0 ng mL–1 selenium, 2 mM glutamine, 2 mM hypoxanthine and 1.25 mg mL–1 bovine serum albumin) in the absence or presence of 200 ng mL–1 GDF-9 and/or 50 ng mL–1 FSH added during the first (Days 0–8) and/or second (Days 8–16) half of the culture period. Non-cultured and cultured fragments were processed for histological and ultrastructural analyses. After 16 days, all treatments using GDF-9 or FSH showed higher rates of follicular survival compared with αMEM+ alone. Compared with non-cultured control, sequential culture media containing GDF-9 and/or FSH significantly increased the percentage of developing follicles and follicle diameter. Moreover, a progressive increase in oocyte diameter was observed only with sequential culture medium containing GDF-9 until Day 8 followed by FSH (GDF-9/FSH) in the second half of the culture period. After 16 days of culture, ultrastructural analysis confirmed the integrity of follicles cultured in the presence of GDF-9/FSH. In conclusion, a dynamic medium containing GDF-9 and FSH (GDF-9/FSH) maintained follicular integrity and promoted activation of primordial follicles and growth during long-term in vitro culture of goat preantral follicles.
26
Lonergan, P., D. Rizos, A. Gutierrez-Adan, et al. "243TEMPORAL DIVERGENCE IN THE PATTERN OF MRNA EXPRESSION IN BOVINE EMBRYOS CULTURED FROM THE ZYGOTE TO BLASTOCYST STAGE IN VITRO OR IN VIVO." Reproduction, Fertility and Development 16, no. 2 (2004): 242. http://dx.doi.org/10.1071/rdv16n1ab243.
Full textAbstract:
The objective of this study was to examine the time during the post-fertilization culture period that gene expression patterns of in vitro cultured bovine embryos diverge from those of their in vivo cultured counterparts. Presumptive bovine zygotes were produced by IVM/IVF of immature oocytes collected from the ovaries of slaughtered animals. At approximately 20h post-insemination (hpi), presumptive zygotes were randomly divided into two culture groups, either in vitro in synthetic oviduct fluid or in vivo, and transferred into the ewe oviduct. Embryos were recovered from both systems at approximately 30hpi (2-cell), two (4-cell), three (8-cell), four (16-cell), five (early morula), six (compact morula) or seven (blastocyst) days pi and snap-frozen for the analysis of transcript abundance using real-time PCR. The transcripts studied were interferon-tau, apoptosis regulator box-a (Bax), connexin 43, sarcosine oxidase, glucose transporter 5, mitochondrial Mn-superoxide dismutase, insulin-like growth factor II, and insulin-like growth factor-I receptor, most of which are known from our previous work to be differentially transcribed in blastocysts derived from culture in vitro or in vivo. Analysis was done on pools of 10 embryos. Data were analyzed using one-way repeated measures ANOVA. The relative abundance of the transcripts studied varied throughout the preimplantation period and was strongly influenced by the culture environment. For example, transcripts for interferon-tau were detected from the 8-cell stage onwards in in vitro-cultured embryos but not until the early morula stage in those cultured in vivo. Levels of this transcript increased significantly at the compact morula and blastocyst stages in both groups but were significantly higher (P&lt;0.05) in in vitro-cultured embryos at both stages. mRNA for Bax was not detected before the 8-cell stage in in vitro cultured embryos and not until the 16-cell stage in in vivo cultured embryos. The abundance of this transcript increased significantly thereafter up to the blastocyst stage in both groups. The level of expression was significantly higher (P&lt;0.05) at all stages of development in in vitro-cultured embryos than those cultured in vivo. The relative abundance of Cx43 transcripts decreased in both in vitro- and in vivo-cultured embryos at the 8- to 16-cell stage. Levels remained low thereafter in the in vitro-cultured embryos but significantly increased in those cultured in vivo. Transcript abundance was significantly higher in in vivo cultured embryos from Day 4 onwards with a ten-fold difference presence at the blastocyst stage. Differences also existed for the other transcripts studied. These data demonstrate that changes in transcript abundance in blastocyst stage embryos are in many cases a consequence of perturbed transcription earlier in development. Depending on the transcript, these differences may be evident in as short as 10h of culture.
27
McKechnie, N. M., M. Boulton, H. L. Robey, F. J. Savage, and I. Grierson. "The cytoskeletal elements of human retinal pigment epithelium: in vitro and in vivo." Journal of Cell Science 91, no. 2 (1988): 303–12. http://dx.doi.org/10.1242/jcs.91.2.303.
Full textAbstract:
The cytoskeletal elements of normal (in situ) and cultured human retinal pigment epithelium (RPE) were studied by a variety of immunocytochemical techniques. Primary antibodies to vimentin and cytokeratins were used. Positive immunoreactivity for vimentin was obtained with in situ and cultured material. The pattern of reactivity obtained with antisera and monoclonals to cytokeratins was more complex. Cytokeratin immunoreactivity could be demonstrated in situ and in cultured cells. The pattern of cytokeratin expression was similar to that of simple or glandular epithelia. A monoclonal antibody that specifically recognizes cytokeratin 18 identified a population of cultured RPE cells that had particularly well-defined filamentous networks within their cytoplasm. Freshly isolated RPE was cytokeratin 18 negative by immunofluorescence, but upon culture cytokeratin 18 positive cells were identifiable. Cytokeratin 18 positive cells were identified in all RPE cultures (other than early primaries), regardless of passage number, age or sex of the donor. In post-confluent cultures cytokeratin 18 cells were identified growing over cytokeratin 18 negative cells, suggesting an association of cytokeratin 18 immunoreactivity with cell proliferation. Immunofluorescence studies of retinal scar tissue from two individuals revealed the presence of numerous cytokeratin 18 positive cells. These findings indicate that RPE cells can be identified by their cytokeratin immunoreactivity and that the overt expression of cytokeratin 18 may be associated with proliferation of human RPE both in vitro and in vivo.
28
Cott, G. R., J. Y. Westcott, and N. F. Voelkel. "Prostaglandin and leukotriene production by alveolar type II cells in vitro." American Journal of Physiology-Lung Cellular and Molecular Physiology 258, no. 4 (1990): L179—L187. http://dx.doi.org/10.1152/ajplung.1990.258.4.l179.
Full textAbstract:
Alveolar type II cells were isolated from adult rats, cultured for 22 h, and individual eicosanoids in the media from unstimulated and stimulated cells were quantified by immunoassay. Stimulation with the calcium ionophore A23187 significantly increased the media levels of prostaglandins (prostaglandin and 6-keto-prostaglandin F1 alpha greater than thromboxane B2). In contrast to previous reports, increased media levels of leukotrienes were also recovered from cells incubated with A23187, but only for cells in culture for less than or equal to 24 h. The production of leukotriene C4 was confirmed by a combination of high-performance liquid chromatography and spectrophotometric analysis. The profile of eicosanoids produced by cultures of alveolar type II cells was distinctly different than that of similarly cultured alveolar macrophages. Finally, stimulation of alveolar type II cell cultures with either a phorbol ester or phospholipase C increased media prostaglandin levels but failed to increase leukotriene levels. We conclude that primary cultures of alveolar type II cells are capable of the de novo metabolism of arachidonic acid to both cyclooxygenase and lipoxygenase products and that the production of leukotrienes is dependent on both time in culture and agonist. Thus alveolar type II cells are a potential source for the production of these eicosanoids in vivo, and the particular lipid mediators produced may vary depending on the pathophysiologic stimulus.
29
Arekar, Ashish R., Janhavi A. Arekar, S. S. Barve, and G. T. Paratkar. "In vitro regeneration of Momordica dioica (Roxb.)." Journal of Applied and Natural Science 4, no. 2 (2012): 297–303. http://dx.doi.org/10.31018/jans.v4i2.268.
Full textAbstract:
Momordica dioica, Roxb. (Family: Cucurbitaceae) commonly called as Kartoli, is an important medicinal plant, which has remained unexplored from the commercial point of view. Considering its scarce availability and the medicinal importance, in vitro cultures were established. Traditionally, M. dioica has been propagated mainly through its tuberous roots and less commonly by seeds. Germination through seeds is very difficult or impossible because of hard seed coat. As an alternative to traditional methods tissue culture offers an efficient method for propagation of M. dioica. Mature seeds were used for the regeneration of M. dioica. The decoated seeds of M. dioica were cultured on Murashige and Skoog basal medium (MS medium) supplemented with various combinations of Auxins (á – naphthaleneacetic acid) and Cytokinins (N6 - benzyl adenine). MS basal medium supplemented with 4.44 µM and 8.88 µM N6 - benzyl adenine (BA) gave rise to maximum number of shoots in 7-8 weeks. In vitro grown shoots were sub cultured on MS medium supplemented with different concentrations of indole-3-butyric acid (IBA) for root initiation. MS medium with 0.049mM indole-3-butyric acid (IBA) showed rooting in 45 days. The regenerated plantlets were successfully hardened in vermiculite.
30
Redel, B. K., L. D. Spate, A. N. Brown, and R. S. Prather. "97 SUPPLEMENTATION WITH FOLATE IN VITRO INCREASES TROPHECTODERM AND TOTAL CELL NUMBER IN IN VITRO-DERIVED PORCINE BLASTOCYSTS." Reproduction, Fertility and Development 24, no. 1 (2012): 161. http://dx.doi.org/10.1071/rdv24n1ab97.
Full textAbstract:
It is vital that improvements are made to current culture environments because in vitro culture systems are suboptimal compared with in vivo. A previous transcriptional profiling endeavour conducted by Bauer et al. (2010 Biol. Reprod. 83, 791–798) identified hundreds of mRNA transcripts that were mis-expressed in porcine embryos fertilized in vivo and then cultured in vitro to Day 6 compared with in vivo Day-6 embryos. Enriched in the downregulated transcripts were 4 genes involved with the one carbon pool by folate KEGG pathway. This downregulation of genes involved with folate metabolism may illustrate an impaired folate homeostasis in embryos cultured in the current culture environment. The objective of this study was to determine the effects folate had on embryo development of in vitro fertilized embryos. Porcine cumulus–oocyte complexes were matured for 44 h in M199 supplemented with epidermal growth factor (EGF), FSH and LH. Oocytes with a visible polar body were selected and fertilized in modified tris buffered medium for 5 h and then placed into porcine zygote medium 3 with 0 mM, 0.2 mM, 0.4 mM and 0.8 mM folate to find the optimal concentration of folate. Twenty-eight hours post-fertilization, cleaved embryos were selected and moved into 25-μL drops of respective culture medium and cultured to Day 6 in a water-saturated atmosphere of 5% CO2, 5% O2, 90% N2, at 38.5°C. To determine the effect folate had on development, the blastocyst rate for each treatment group was measured. Results were log-transformed and analysed by using PROC GLM in SAS (SAS Institute Inc., Cary, NC). A least-significant difference post-test comparison was completed to determine if significant differences existed between treatment groups. The percentage of cleaved embryos on Day 6 that developed to blastocyst was 56.2%, 55.9%, 66.9% and 61.8% (n = 133, 149, 135 and 135) in 0 mM, 0.2 mM folate, 0.4 mM folate and 0.8 mM, respectively. The 0.4 mM folate group tended (P = 0.07) to have a higher number of cleaved embryos that developed to the blastocyst stage. Consequently, this concentration was used for all further embryo culture experiments. Differential staining was completed to compare the number of trophectoderm and inner cell mass nuclei for embryos cultured in 0 mM or 0.4 mM folate concentrations. Staining revealed that embryos cultured with folate had an increase in number of trophectoderm (29.7 ± 1.5 vs 24.4 ± 1.4 cells; P = 0.0058) and total cell (36.9 ± 1.0 vs 31.7 ± 1.0; P = 0.0007) numbers compared with embryos cultured without folate. These results illustrate that the addition of folate to current culture medium doesn't hinder development to blastocyst and by increasing trophectoderm and total cell number may give rise to better-quality in vitro-derived embryos. It is evident that using transcriptional profiling can be a great method of identifying ways to improve embryo culture systems and, in this case, supplementing with folate. Funded by Food for the 21st Century.
31
Edelman, P. D., D. C. McFarland, V. A. Mironov, and J. G. Matheny. "Commentary: In Vitro-Cultured Meat Production." Tissue Engineering 11, no. 5-6 (2005): 659–62. http://dx.doi.org/10.1089/ten.2005.11.659.
Full text
32
Teryukova, N. P., G. I. Blinova, and V. A. Ivanov. "Zajdela hepatoma cells cultured in vitro." Cell and Tissue Biology 7, no. 3 (2013): 245–52. http://dx.doi.org/10.1134/s1990519x13030127.
Full text
33
Marín, M. L., and J. A. Marín. "Excised rootstock roots cultured in vitro." Plant Cell Reports 18, no. 3-4 (1998): 350–55. http://dx.doi.org/10.1007/s002990050585.
Full text
34
Sluss, Patrick M., Kan-ei Lee, John H. Mattox, Patricia C. Smith, Margaret C. Graham, and Ann B. Partridge. "Estradiol and progesterone production by cultured granulosa cells cryopreserved from in vitro fertilization patients." European Journal of Endocrinology 130, no. 3 (1994): 259–64. http://dx.doi.org/10.1530/eje.0.1300259.
Full textAbstract:
Sluss PM, Lee K, Mattox JH, Smith PC, Graham MC, Partridge AB. Estradiol and progesterone production by cultured granulosa cells cryopreserved from in vitro fertilization patients. Eur J Endocrinol 1994;130:259–64. ISSN 0804–4643 Gonadotropin-stimulated steroidogenesis was studied in cultured human granulosa-lutein cells obtained from patients undergoing procedures for in vitro fertilization. The impact of cryopreservation on cell function in vitro was studied. Granulosa cells obtained from in vitro fertilization patients were cultured in serum-supplemented medium or cryopreserved at −135°C for 2–22 months. Fresh (unfrozen) cells (105) produced estradiol at a rate of 1320 pmol/l (over 72 h) and progesterone at about 2500 nmol/l. Estradiol production by either fresh or cryopreserved granulosa cells in culture was unaffected by physiological concentrations of follicle-stimulating hormone (7IU/l). Adding testosterone (10−7 mol/l) to the medium increased estradiol secretion approximately sixfold. In contrast, progesterone production was not affected by follicle-stimulating hormone or testosterone. No significant differences were observed in cultures of cryopreserved granulosa cells compared to cultures of unfrozen cells with respect to estradiol secretion, the effects of follicle-stimulating hormone or testosterone on estradiol secretion, or progesterone production. Progesterone production by fresh and cryopreserved cells was stimulated by human chorionic gonadotropin. These data indicate that cryopreservation offers the potential to facilitate prospective studies utilizing large numbers of human granulosa-lutein cells in culture. Patrick M Sluss, Reproductive Endocrine Unit, Massachusetts General Hospital East, 149 13th Street, Charlestown, MA 02129, USA
35
Silva, T. F., S. L. Costa, E. P. Costa, J. D. Guimarães, and V. L. D. Queiroz-Castro. "Effect of somatotropin on survival and diameter of bovine preantral follicles." Arquivo Brasileiro de Medicina Veterinária e Zootecnia 71, no. 5 (2019): 1445–52. http://dx.doi.org/10.1590/1678-4162-10602.
Full textAbstract:
ABSTRACT The aim of this study was to evaluate the effect of Recombinant bovine somatotropin (rbST) on survival and diameter of bovine preantral ovarian follicles (PAOF) cultured in vitro. Ovaries were collected from adult cows and fragments of ovarian cortex were immediately fixed (non-cultured control) or cultured in vitro in α-MEM+ alone or containing 10, 50, 100 or 1,000ng/mL rbST. The fragments were processed for Classical Histology and Transmission Electron Microscopy. After one and seven days of culture, the percentage of normal follicles in the non-cultured control was superior (P< 0.05) to the follicles cultured in α-MEM+ alone or with different rbST concentrations. The oocyte and follicular mean diameter did not increase during the culture for one and seven days, both in media containing rbST and in the medium without this hormone. The only medium in which there was no reduction in follicular diameter with the time of culture was the medium without rbST. Ultrastructural damage in PAOF cultured in vitro was found. It is concluded that the use of rbST at different concentrations in in situ culture of bovine preantral follicles has no beneficial effects on survival and growth of bovine PAOF.
36
RANKIN, KENNETH S., RACHEL L. LAKEY, CRAIG H. GERRAND, ANDREW P. SPROWSON, ANDREW W. McCASKIE, and MARK A. BIRCH. "A Novel in Vitro Model to Investigate Behavior of Articular Chondrocytes in Osteoarthritis." Journal of Rheumatology 37, no. 2 (2009): 426–31. http://dx.doi.org/10.3899/jrheum.080080.
Full textAbstract:
Objective. To investigate in vivo simulation of the microenvironment in which osteoarthritis (OA) chondrocytes are cultured in vitro.Methods. Human articular chondrocytes were cultured under normoxic and hypoxic conditions. Cells were cultured on standard culture plastic or a porous polyHEMA surface that closely resembles the in vivo cartilage microarchitecture. Morphological changes to the cells were demonstrated by fluorescent staining with DAPI and vinculin. Proteoglycan and type II collagen protein levels were assessed using established techniques. Matrix metalloproteinase-1 (MMP-1) production was assessed by ELISA. The gene expression of type II collagen and SOX9 was measured using real-time polymerase chain reaction.Results. Cells grown on culture plastic were seen to be flat and hexagonal. Cells cultured on the porous polyHEMA surface exhibited morphology in keeping with the in vivo microenvironment. Glycosaminoglycan release in hypoxia was high from cells cultured on standard culture plastic. Transcriptional expression of type II collagen was upregulated in hypoxia and by culture on the polyHEMA surface. Transcriptional expression of SOX9 in hypoxia was upregulated compared to normoxia; no significant effect was seen by varying the culture surface. Translational expression of type II collagen was upregulated at 20% oxygen on the polyHEMA surface compared to culture plastic and this was related to MMP-1 expression.Conclusion. Culture of chondrocytes in hypoxia and on a porous surface simulates the in vivo microenvironment and illustrates the molecular mechanisms of OA.
37
Li, Ming, Yong-Hai Li, Yi Hou, Xiao-Fang Sun, Qingyuan Sun, and Wei-Hua Wang. "Isolation and culture of pluripotent cells from in vitro produced porcine embryos." Zygote 12, no. 1 (2004): 43–48. http://dx.doi.org/10.1017/s0967199404002679.
Full textAbstract:
The present study was designed to examine whether in vitro produced porcine embryos can be used to establish an embryonic stem (ES) cell line. Porcine embryos were produced by in vitro maturation and in vitro fertilization. Embryos at the 4-cell to blastocyst stages were cultured in an ES medium containing 16% fetal bovine serum with mouse embryonic fibroblasts as a feeder layer. It was found that ES-like colonies were derived only from blastocysts. When these ES-like colonies were separated in 0.25% trypsin–0.02% EDTA solution and cultured again, ES-like colonies were further observed in the subsequent culture until the fourth passage. The cells from ES-like colonies showed positive alkaline phosphatase activity. Some cells from the colonies differentiated into several types of cells in vitro when they were cultured in the medium without feeder layers and leukemin inhibitory factor. Embryoid bodies were also formed when the cells were cultured in a suspension status. These results indicate that porcine ES-like cells can be derived from in vitro produced porcine blastocysts and these ES-like cells are pluripotent. The culture system used in the present study is useful to isolate and culture ES cells from in vitro produced porcine embryos.
38
Gupta, P. S. P., H. S. Ramesh, B. M. Manjunatha, S. Nandi, and J. P. Ravindra. "Production of buffalo embryos using oocytes from in vitro grown preantral follicles." Zygote 16, no. 1 (2008): 57–63. http://dx.doi.org/10.1017/s096719940700442x.
Full textAbstract:
SummaryThe present study examines the use of buffalo preantral follicles as a source of oocytes for in vitro embryo production. Preantral follicles were isolated from abattoir-derived buffalo ovaries and were grown for 100 days in five different culture systems: (1) minimum essential medium (MEM); (2) coconut water; (3) MEM + ovarian mesenchymal cell (OMC) co-culture; (4) MEM + granulosa cell (GC) co-culture; or (5) MEM + cumulus cell (CC) co-culture. Low growth rates for the preantral follicles were observed when follicles were cultured in MEM or coconut water medium. Moderate growth rates were seen for OMC and GC co-cultures, and high rates of growth were observed when follicles were grown in CC co-culture. The survival of preantral follicles was low in the MEM culture (<25%), but was over 75% in the other culture systems. Oocytes were not recovered from the MEM group, while an oocyte recovery rate of 80–100% was observed when the follicles were cultured with coconut water/somatic cells. Transferable embryos could be produced only with the oocytes obtained from preantral follicles grown in the OMC and CC co-culture systems. This study demonstrates, for the first time, that it is possible to produce buffalo embryos by in vitro fertilization of oocytes derived from in vitro grown preantral follicles.
39
Buchala, A. J., T. Genoud, and H. Meier. "Polysaccharides in the culture medium of cotton cells cultured in vitro." Food Hydrocolloids 1, no. 5-6 (1987): 359–63. http://dx.doi.org/10.1016/s0268-005x(87)80026-7.
Full text
40
Carrara, Maria, Lorenzo Cima, Roberto Cerini, and Maurizio Dalle Carbonare. "A New In Vitro Model for Predicting the Toxicity of Cosmetic Products." Alternatives to Laboratory Animals 20, no. 1 (1992): 138–43. http://dx.doi.org/10.1177/026119299202000119.
Full textAbstract:
A method has been developed whereby cosmetic products which are not soluble in water or in alcohol can be brought into contact with cell cultures by being placed in a cell culture insert, which is then placed in the cell culture well. Preliminary experiments were carried out with L929 cells, and cytotoxicity was evaluated by measuring neutral red uptake and the total protein content of treated cultured cells. Encouraging results were obtained in comparisons of three cosmetic emulsions and of one emulsion containing a range of concentrations of two preservatives, Kathon CG and Bronopol.
41
Villalobos, Victor M., Edward C. Yeung, and Trevor A. Thorpe. "Origin of adventitious shoots in excised radiata pine cotyledons cultured in vitro." Canadian Journal of Botany 63, no. 12 (1985): 2172–76. http://dx.doi.org/10.1139/b85-307.
Full textAbstract:
Previous studies on shoot initiation in cultured excised cotyledons of Pinus radiata indicated that the process began early in culture on the side of the cotyledon in contact with the cytokinin-containing medium. In contrast to cotyledons cultured without cytokinin, organized structures, which have been termed promeristemoids, can be observed in cotyledons cultured with cytokinin after 5 days in culture. These structures arise from single cells in the first subepidermal cell layer at day 3. They are stable and their continued division leads to the formation of meristemoids, shoot primordia, and finally shoots with primary needles.
42
Schinor, Evandro Henrique, Fernando Alves de Azevedo, Francisco de Assis Alves Mourão Filho, and Beatriz Madalena Januzzi Mendes. "In vitro organogenesis in some citrus species." Revista Brasileira de Fruticultura 33, no. 2 (2011): 526–31. http://dx.doi.org/10.1590/s0100-29452011005000050.
Full textAbstract:
In vitro organogenesis of Citrus was studied for the genotypes Citrus sinensis cv. 'Natal', C. limonia, C. volkameriana, and C. aurantium, with the use of epicotyl segments-derived explants, cultured in MT salts and vitamins medium supplemented with different concentrations of 6-benzylaminopurine (BAP - 0.0; 0.5; 1.0; 1.5 or 2.0 mg L-1). For the recalcitrant genotypes C. limonia and C. aurantium the in vitro organogenesis was also studied with internodal segments-derived explants, cultured in MT salts and vitamins medium supplemented with 0; 0.5; 1.0; 2.0, or 4.0 mg L-1 of BAP. The efficiency of culture medium supplementation with the combination of BAP (0.0; 1.0, or 2.0 mg L-1) and NAA (1-naphthaleneacetic acid - 0.0; 0.3, or 0.5 mg L-1) in the development of adventitious shoots was evaluated for C. aurantium. Culture medium supplementation with BAP is not essential for the adventitious shoots development in the four genotypes studied when epicotyl segments-derived explants are used. In general, culture media supplementation with BAP decreased the percentage of responsive explants excepted for C. sinensis cv. 'Natal' and C. limonia when the concentrations of 1.5 and 2.0 mg/L were used. The presence of cytokinin, in concentrations up to 2 mg/L, stimulated the in vitro organogenesis when internodal segments-derived explants were used for C. limonia and C. aurantium. For C. aurantium no adventitious shoots developed in explants (internodal segments) cultured in basal culture medium, without BAP supplementation. Although no statistic differences could be detected, culture media supplementation with the combination of BAP and NAA favored the development of adventitious shoots in C. aurantium. The best concentration of NAA varied according to BAP concentration. The results presented herein, show that Citrus in vitro organogenesis depends on the interaction of culture medium composition, explant differentiation level, and genotype.
43
Lima-Verde, I. B., M. H. T. Matos, J. B. Bruno та ін. "Effects of α-tocopherol and ternatin antioxidants on morphology and activation of goat preantral follicles in vitro cultured". Arquivo Brasileiro de Medicina Veterinária e Zootecnia 61, № 1 (2009): 57–65. http://dx.doi.org/10.1590/s0102-09352009000100009.
Full textAbstract:
The effects of α-tocopherol and ternatin on the morphology, activation, and growth of goat preantral follicles in vitro cultured, for one or five days, were evaluated. Ovarian fragments were immediately fixed (non-cultured control) or in vitro cultured for one or five days in Minimum Essential Medium (MEM) with or without α-tocopherol or ternatin supplementation, both at concentrations of 5, 10, or 15µM, corresponding to the following treatments: MEM, TOC5, TOC10, TOC 15, TER5, TER10, and TER15. The percentages of morphologically normal preantral follicles in non-cultured ovarian tissue (control) was 73.2% and after five days of culture, there was a decrease on these percentages in all treatments (P<0.05) when compared with non-cultured control. Culture of ovarian cortex for five days increased the percentages of follicular activation in all treatments (P<0.05). Ultrastructural analysis did not confirm the integrity of caprine preantral follicles cultured for five days in medium containing antioxidants. This study demonstrated that α-tocopherol and ternatin can promote follicular activation; however, addition of these antioxidants in the tested concentrations reduced the follicular viability after in vitro culture.
44
Pizza, Francis X., Thomas J. McLoughlin, Stephen J. McGregor, Edward P. Calomeni, and William T. Gunning. "Neutrophils injure cultured skeletal myotubes." American Journal of Physiology-Cell Physiology 281, no. 1 (2001): C335—C341. http://dx.doi.org/10.1152/ajpcell.2001.281.1.c335.
Full textAbstract:
The purpose of the study was to test the hypothesis that neutrophils can injure cultured skeletal myotubes. Human myotubes were grown and then cultured with human blood neutrophils. Myotube injury was quantitatively and qualitatively determined using a cytotoxicity (51Cr) assay and electron microscopy, respectively. For the 51Cr assay, neutrophils, under non-in vitro-stimulated and N-formylmethionyl-leucyl-phenylalanine (FMLP)-stimulated conditions, were cultured with myotubes at effector-to-target cell (E:T) ratios of 10, 30, and 50 for 6 h. Statistical analyses revealed that myotube injury was proportional to the E:T ratio and was greater in FMLP-stimulated conditions relative to non-in vitro-stimulated conditions. Transmission electron microscopy, using lanthanum as an extracellular tracer, revealed in cocultures a diffuse appearance of lanthanum in the cytoplasm of myotubes and a localized appearance within cytoplasmic vacuoles of myotubes. These observations and their absence in control cultures (myotubes only) suggest that neutrophils caused membrane rupture and increased myotube endocytosis, respectively. Myotube membrane blebs were prevalent in scanning and transmission electron micrographs of cultures consisting of neutrophils and myotubes (E:T ratio of 5) and were absent in control cultures. These data support the hypothesis that neutrophils can injure skeletal myotubes in vitro and may indicate that neutrophils exacerbate muscle injury and/or delay muscle regeneration in vivo.
45
Pellegrini, Graziella, Roberto Gherzi, Adriano Tito Franzi, Fiorella D'Anna, Michele De Luca, and Ranieri Cancedda. "In Vitro Reconstitution of Differentiated Human Epithelia." Alternatives to Laboratory Animals 20, no. 1 (1992): 95–102. http://dx.doi.org/10.1177/026119299202000113.
Full textAbstract:
Human keratinocytes obtained from skin biopsies can be serially cultured in vitro. When plated on lethally irradiated 3T3 fibroblasts, keratinocyte colonies reconstitute a stratified squamous epithelium devoid of stratum corneum. The expression of a mature cornified epidermis, expressing all the morphological and biochemical markers of the in vivo epidermis, can be obtained by the “emerged dermal equivalent” culture system. Melanocytes grown under the same culture conditions, maintain a physiological melanocyte/keratinocyte ratio, are organised in the basal layer of the cultured epidermis, and maintain differentiated functions such as dendritic arborisation, melanin synthesis and melanosome transfer. This allows the reconstitution of an epidermis physiologically populated by functionally active melanocytes. Epithelial cells from different mucosal body sites, namely palate, urethra, conjunctiva, cornea and vagina, can also be cultured and maintain the characteristics of the original donor sites. The in vitro reconstituted human epithelia, permanently transplanted onto patients presenting large epidermal or mucosal defects, retain the characteristics of the original donor site, suggesting an intrinsic site-specific differentiation programme. These three-dimensional human epithelium models could prove useful in standard cytotoxicity assays and could be used as a tool to study the effects of a variety of compounds on normal human epithelia in vitro.
46
Jia, Zhidong, Yuan Cheng, Xinan Jiang, et al. "3D Culture System for Liver Tissue Mimicking Hepatic Plates for Improvement of Human Hepatocyte (C3A) Function and Polarity." BioMed Research International 2020 (March 4, 2020): 1–22. http://dx.doi.org/10.1155/2020/6354183.
Full textAbstract:
In vitro 3D hepatocyte culture constitutes a core aspect of liver tissue engineering. However, conventional 3D cultures are unable to maintain hepatocyte polarity, functional phenotype, or viability. Here, we employed microfluidic chip technology combined with natural alginate hydrogels to construct 3D liver tissues mimicking hepatic plates. We comprehensively evaluated cultured hepatocyte viability, function, and polarity. Transcriptome sequencing was used to analyze changes in hepatocyte polarity pathways. The data indicate that, as culture duration increases, the viability, function, polarity, mRNA expression, and ultrastructure of the hepatic plate mimetic 3D hepatocytes are enhanced. Furthermore, hepatic plate mimetic 3D cultures can promote changes in the bile secretion pathway via effector mechanisms associated with nuclear receptors, bile uptake, and efflux transporters. This study provides a scientific basis and strong evidence for the physiological structures of bionic livers prepared using 3D cultures. The systems and cultured liver tissues described here may serve as a better in vitro 3D culture platform and basic unit for varied applications, including drug development, hepatocyte polarity research, bioartificial liver bioreactor design, and tissue and organ construction for liver tissue engineering or cholestatic liver injury.
47
Mercer, J. G., A. E. Munn, J. W. Smith, and H. H. Rees. "Cuticle production and ecdysis in larval marine ascaridoid nematodes in vitro." Parasitology 92, no. 3 (1986): 711–20. http://dx.doi.org/10.1017/s0031182000065562.
Full textAbstract:
SUMMARYInfective 3rd-stage larvae of three species of marine ascaridoid nematodes, Pseudoterranova (= Phocanema = Porrocaecum = Terranova) decipiens, Anisakis simplex and Phocascaris/Contracaecum sp. were cultured in vitro in 0·9% saline or Medium 199 at 37°C beneath an atmosphere of either air or 95% air:5% CO2 All three species responded to culture at 37°C by producing a new cuticle. The majority of P. decipiens completed ecdysis under all the culture conditions employed. Ecdysis in A. simplex was stimulated by a high concentration of CO2; this effect was particularly noticeable in saline cultures. Both A. simplex and Phocascaris/Contracaecum sp. developed more effectively in a nutrient medium than in saline.
48
Yamauchi, N., H. Sasada, S. Sugawara, and T. Nagai. "Effect of culture conditions on artificial activation of porcine oocytes matured in vitro." Reproduction, Fertility and Development 8, no. 8 (1996): 1153. http://dx.doi.org/10.1071/rd9961153.
Full textAbstract:
The effects of culture media used and culture period for in vitro maturation of porcine oocytes on their subsequent response to chemical and electrical activation, were investigated. Activated oocytes were identified by the presence of a pronucleus(ei) or cleavage. Porcine oocytes were cultured for 24, 30, 36, 42 and 48 h in TCM199 with Earle's salts (199) supplemented with 10% fetal calf serum (199-FCS) before electrical stimulation. Although few oocytes were activated after 24 h and 30 h of culture (5.4% and 6.1% respectively), the percentage of activated oocytes increased significantly to 93.2% after 42 h in culture (P < 0.05); however, when the culture period was extended to 48 h, there was a significant decrease to 56.7% (P < 0.05). Oocytes were also cultured in four types of media: (1) 199-FCS; (2) 199 supplemented with 5 mg mL-1 bovine serum albumin (199-BSA); (3) Kreb's-Ringer bicarbonate solution supplemented with 10% FCS (KRB-FCS); and (4) KRB supplemented with BSA (KRB-BSA). After 42 h of culture in each medium, the oocytes were electrically activated. Although rates of maturation of oocytes cultured in the four media were similar (74.0-80.8%), all oocytes except those cultured in 199-FCS failed to be activated. In addition, oocytes were cultured for 36, 42 and 48 h in 199-FCS and then stimulated by treatment with ethanol. Significantly fewer oocytes were activated in the chemically-treated group than in the electrically-treated group. These results indicate that culture conditions used for the culture of porcine oocytes in vitro are important with respect to their subsequent response to artificial activation.
49
Adelberg, J., M. Kroggel, and J. Toler. "Physical Environment in vitro Affects Laboratory and Nursery Growth of Micropropagated Hostas." HortTechnology 10, no. 4 (2000): 754–57. http://dx.doi.org/10.21273/horttech.10.4.754.
Full textAbstract:
Hosta ×hybrid Tratt. `Blue Cadet' and Hosta tokudama Tratt. `Newberry Gold' were micropropagated in shaken liquid culture and on agar media, in conventional vessels and vessels modified for ventilation in vitro. Acclimatization under intermittent mist and growth in an outdoor nursery during the late spring and summer were monitored by dry weight analysis of sample plants every 4 days for a 60-day period (ex vitro growth). Results for `Newberry Gold' were 1) in vitro shoot growth was greater in liquid than agar culture, regardless of vessel; 2) shoots from agar or liquid culture grew at similar rates ex vitro; 3) ex vitro root growth was greater for liquid than agar cultured plants, regardless of vessel type. Results for `Blue Cadet' were 1) in vitro and ex vitro shoot growth was greater in liquid than agar culture regardless of vessel type and 2) ex vitro root growth was greatest for liquid cultured plants from conventional vessels. Ventilated vessels were generally beneficial for agar but not liquid culture. Benefits of liquid culture for micropropagation of Hosta found in vitro are at least maintained and sometimes enhanced during ex vitro growth in the mist bed and nursery.
50
Tablin, F., M. Castro, and R. M. Leven. "Blood platelet formation in vitro. The role of the cytoskeleton in megakaryocyte fragmentation." Journal of Cell Science 97, no. 1 (1990): 59–70. http://dx.doi.org/10.1242/jcs.97.1.59.
Full textAbstract:
We have developed a unique in vitro model that promotes differentiation of megakaryocytes into platelets. When megakaryocytes isolated from guinea pig bone marrow were cultured on hydrated rat tail collagen gels, cells spontaneously formed elongated, beaded processes that fragmented to yield cytoplasmic pieces with the same size and internal composition as individual platelets. Addition of nocodazole at the initiation of cultures blocked process formation, while addition of nocodazole to cells with previously established processes resulted in their retraction. The addition of taxol to cultures resulted in abnormally thick processes that were tightly adherent to the underlying substratum, and did not bead or fragment. Cytochalasin D accelerated process formation and fragmentation of megakaryocytes cultured on collagen gels by twofold. On the basis of these results, we propose a model for platelet formation in culture that involves the following steps: adherence of megakaryocytes to the underlying extracellular matrix; dilation of the demarcation membrane system and breakdown of the actin-rich peripheral zone; microtubule-based extension of pseudopodia, which are no longer adherent to the substratum; and fragmentation into platelets by the coalescence and fusion of demarcation membrane vesicles with the plasma membrane. We feel that this distinctive culture system closely approximates thrombocytopoiesis in vivo, thus allowing detailed elucidation of this important process.