Relevant bibliographies by topics / Cultures Cells

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  1. Journal articles
  2. Dissertations / Theses
  3. Books
  4. Book chapters
  5. Conference papers
  6. Reports

Academic literature on the topic 'Cultures Cells'

Author: Grafiati
Published: 4 June 2021
Last updated: 14 February 2022

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Journal articles on the topic "Cultures Cells"

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First, NL, MM Sims, SP Park, and MJ Kent-First. "Systems for production of calves from cultured bovine embryonic cells." Reproduction, Fertility and Development 6, no. 5 (1994): 553. http://dx.doi.org/10.1071/rd9940553.

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The development of totipotent bovine embryonic cell cultures has great value in cattle breeding. They provide: (1) a mechanism for making large numbers of clonal offspring by nuclear transfer; (2) an efficient gene transfer system through the use of selectable markers to select transgenic cells; and (3) a mechanism for site-specific gene transfer or deletion by homologous DNA sequence recombination. Bovine embryonic cell cultures have been established from blastocyst inner cell mass (ICM) cells, morulae and the precompaction 16-20-cell stage. All have exhibited similar morphology to mouse embryonic stem (ES) cells, pluripotency on differentiation and proliferation in culture. Culture systems have consisted of microdrop loose suspension short-term cultures or long-term cultures on bovine or murine fibroblast feeder layers, in either a microdrop or a culture dish. The relative merit of culture systems or media requirements for mitosis and prevention of differentiation have not been determined. At present, totipotency is also unknown for cultured cells of the 16-20-cell stage. For cultured ICM cells, totipotency was demonstrated by the birth of four calves from ICM cells cultured 27 days or less in a loose suspension microdrop. Advanced pluripotency and perhaps totipotency was demonstrated in one fetus in a recently reported study where morulae cells cultured in vitro were chimaerized with non-cultured cells. DNA fingerprinting to associate cell lines with offspring and karyotyping to ascertain chromatin normalcy is important in ES cell research. Data pertaining to the use of each are presented.
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Huang, Xiaosong, L. Jeanne Pierce, Paul A. Cobine, Dennis R. Winge, and Gerald J. Spangrude. "Copper Modulates the Differentiation of Mouse Hematopoietic Progenitor Cells in Culture." Cell Transplantation 18, no. 8 (August 2009): 887–97. http://dx.doi.org/10.3727/096368909x471152.

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Copper chelation has been shown to favor the expansion of human hematopoietic stem/progenitor cells in vitro. To further understand the effects of copper modulation on defined subsets of stem cells versus progenitor cells, we extended the studies in a mouse system. We isolated mouse hematopoietic stem cells (HSCs) or hematopoietic progenitor cells (HPCs) and cultured them with or without the copper chelator tetraethylenepentamine (TEPA) or CuCl2. Cytokine-stimulated HPC cultures treated with TEPA for 7 days generated about two to three times more total and erythroid colony-forming cells (CFCs) compared to control cultures. In contrast, CuCl2 treatment decreased the CFC numbers. Similar results were seen with HSC after 14, but not 7, days of culture. Transplant studies showed that HPCs cultured for 7 days in TEPA had about twofold higher short-term erythroid repopulation potential compared to control cultures, while CuCl2 decreased the erythroid potential of cultured HPCs compared to control cultures. HSCs cultured with TEPA for 7 days did not exhibit significantly higher repopulation potential in either leukocyte or erythrocyte lineages compared to control cultures in short-term or long-term assays. Based on JC-1 staining, the mitochondrial membrane potential of HPCs cultured with TEPA was lower relative to control cultures. Our data suggest that decreasing the cellular copper content with TEPA results in preferential expansion or maintenance of HPC that are biased for erythroid differentiation in vivo, but does not enhance the maintenance of HSC activity in culture.
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Agius, L. "Metabolic interactions of parenchymal hepatocytes and dividing epithelial cells in co-culture." Biochemical Journal 252, no. 1 (May 15, 1988): 23–28. http://dx.doi.org/10.1042/bj2520023.

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When parenchymal hepatocytes isolated from adult liver are co-cultured with other epithelial cells, the production of various plasma proteins by the hepatocytes is preserved for much longer than in conventional culture. This study examines some of the metabolic interactions between parenchymal hepatocytes and epithelial cells maintained in co-culture. The leakage of lactate dehydrogenase by hepatocytes co-cultured with epithelial cells was lower than in conventional hepatocyte culture. The epithelial cells have a high glycolytic rate and provide the hepatocytes with a continual supply of lactate. The [lactate] was lower in co-cultures of hepatocytes and epithelial cells than in pure epithelial cultures of similar density, suggesting lactate clearance by the hepatocytes. Alanine uptake was higher in conventional hepatocyte cultures, which lack an exogenous supply of lactate, than in parenchymal hepatocytes in co-culture. Studies with pure parenchymal hepatocytes incubated with increasing [lactate] suggest that lactate is utilized in preference to alanine as a gluconeogenic substrate by hepatocytes co-cultured with epithelial cells. Ketogenesis and carnitine palmitoyltransferase activity declined more slowly in hepatocytes co-cultured with epithelial cells than in conventional culture. It is concluded that the co-culture model has potential for long-term studies of carbohydrate and lipid metabolism.
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Sandstrom, CE, JG Bender, ET Papoutsakis, and WM Miller. "Effects of CD34+ cell selection and perfusion on ex vivo expansion of peripheral blood mononuclear cells." Blood 86, no. 3 (August 1, 1995): 958–70. http://dx.doi.org/10.1182/blood.v86.3.958.958.

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Abstract Ex vivo expansion of peripheral blood mononuclear cells (MNCs), cultured both directly and after selection for CD34+ cells, was compared in static and continuously perfused cultures containing interleukin (IL)-3, IL-6, granulocyte colony-stimulating factor (G- CSF), and stem cell factor (SCF). Cultures inoculated with either MNCs or CD34+ cells produced cells that were remarkably similar after 10 days of culture, as evidence by cell morphology, expression of CD34, CD33, CD15, and CD11b, and the fractions of cells giving rise to colony- forming units granulocyte-monocyte (CFU-GM) and long-term culture- initiating cells (LTC-IC). Static and perfusion cultures gave similar average total cells and CFU-GM expansions for both MNC and CD34+ cell cultures. However, those samples that performed poorly in static culture performed at near-normal levels in perfusion. In addition, perfusion supported higher LTC-IC numbers for both MNC and CD34+ cell cultures. While total cell expansion was about ten times greater in CD34+ cell cultures (approximately 100-fold), CFU-GM expansion (approximately 20-fold) was similar for both MNC and CD34+ cell cultures. The similar distribution of cell types produced in MNC and CD34+ cell cultures allows direct comparison of total and colony- forming cell production. After 15 days in perfusion, MNC cultures produced 1.5-, 2.6-, and 2.1-fold more total cells, CFU-GM, and LTC-IC, respectively, than the same sample selected and cultured as CD34+ cells. Even if the CD34+ selection process was 100% efficient, CFU-GM production would be 1.5-fold greater for MNCs than for CD34+ cells.
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Sandstrom, CE, JG Bender, ET Papoutsakis, and WM Miller. "Effects of CD34+ cell selection and perfusion on ex vivo expansion of peripheral blood mononuclear cells." Blood 86, no. 3 (August 1, 1995): 958–70. http://dx.doi.org/10.1182/blood.v86.3.958.bloodjournal863958.

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Ex vivo expansion of peripheral blood mononuclear cells (MNCs), cultured both directly and after selection for CD34+ cells, was compared in static and continuously perfused cultures containing interleukin (IL)-3, IL-6, granulocyte colony-stimulating factor (G- CSF), and stem cell factor (SCF). Cultures inoculated with either MNCs or CD34+ cells produced cells that were remarkably similar after 10 days of culture, as evidence by cell morphology, expression of CD34, CD33, CD15, and CD11b, and the fractions of cells giving rise to colony- forming units granulocyte-monocyte (CFU-GM) and long-term culture- initiating cells (LTC-IC). Static and perfusion cultures gave similar average total cells and CFU-GM expansions for both MNC and CD34+ cell cultures. However, those samples that performed poorly in static culture performed at near-normal levels in perfusion. In addition, perfusion supported higher LTC-IC numbers for both MNC and CD34+ cell cultures. While total cell expansion was about ten times greater in CD34+ cell cultures (approximately 100-fold), CFU-GM expansion (approximately 20-fold) was similar for both MNC and CD34+ cell cultures. The similar distribution of cell types produced in MNC and CD34+ cell cultures allows direct comparison of total and colony- forming cell production. After 15 days in perfusion, MNC cultures produced 1.5-, 2.6-, and 2.1-fold more total cells, CFU-GM, and LTC-IC, respectively, than the same sample selected and cultured as CD34+ cells. Even if the CD34+ selection process was 100% efficient, CFU-GM production would be 1.5-fold greater for MNCs than for CD34+ cells.
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Eswaramoorthy, Sindhuja D., Nandini Dhiman, Gayathri Korra, Carlo M. Oranges, Dirk J. Schaefer, Subha N. Rath, and Srinivas Madduri. "Isogenic-induced endothelial cells enhance osteogenic differentiation of mesenchymal stem cells on silk fibroin scaffold." Regenerative Medicine 14, no. 7 (July 2019): 647–61. http://dx.doi.org/10.2217/rme-2018-0166.

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Aim: We investigated the role of induced endothelial cells (iECs) in mesenchymal stem cells (MSCs)/iECs co-culture and assessed their osteogenic ability on silk fibroin nanofiber scaffolds. Methods: The osteogenic differentiation was assessed by the ALP assay, calcium assay and gene expression studies. Results: The osteogenic differentiation of the iECs co-cultures was found to be higher than the MSCs group and proximal to endothelial cells (ECs) co-cultures. Furthermore, the usage of isogenic iECs for co-culture increased the osteogenic and endothelial gene expression. Conclusion: These findings suggest that iECs mimic endothelial cells when co-cultured with MSCs and that one MSCs source can be used to give rise to both MSCs and iECs. The isogenic MSCs/iECs co-culture provides a new option for bone tissue engineering applications.
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Stano, J., K. Mičieta, E. Tokhtaeva, M. Valšíková, M. Koreňová, and V. Blanáriková. "Demonstration of lactase activity in culture medium of melon cells." Horticultural Science 31, No. 4 (November 25, 2011): 132–35. http://dx.doi.org/10.17221/3806-hortsci.

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Lactase activity was detected in a culture medium of the cell suspension culture of watermelon (Citrullus vulgaris L.). A simple, rapid and reproducible procedure for identification of extracellular lactase is described using callus cultures of seedlings from the tested plant, hairy roots of 2.5 days old seedlings of watermelon germinating on agar plates as well as cell suspension cultures derived from callus cultures. For the determination of intracellular activities of lactase, 6-bromo-2-naphthyl-β-D-galactopyranoside and p-nitrophenyl-β-D-galactopyranoside were used as synthetic substrates. The extracellular lactase activity was determined by evaluating the day-zone in agar medium. The enzyme from watermelon callus cultures and seedling roots, cultivated on agar plates supplemented with 6-bromo-2-naphthyl-2-bromo-β-D-galactopyranoside, hydrolyzed this substrate releasing 6-bromo-naphthyl. By simultaneous coupling with hexazonium p-rosaniline or Fast Blue BB the corresponding azo dye was formed. The parallel extracellular and intracellular activities were determined in cell suspension cultures derived from callus cultures. The results show a 43.8% intracellular and 54.2% extracellular distribution of lactase activity. The described agar plate method enables a rapid, simple and specific detection of plant processes of extracellular lactase.  
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Culp, D. J., and L. R. Latchney. "Mucinlike glycoproteins from cat tracheal gland cells in primary culture." American Journal of Physiology-Lung Cellular and Molecular Physiology 265, no. 3 (September 1, 1993): L260—L269. http://dx.doi.org/10.1152/ajplung.1993.265.3.l260.

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In a recent study (D. J. Culp, D. K. P. Lee, D. P. Penney, and M. G. Marin. Am. J. Physiol. 263: L264-275, 1992), we reported that primary cultures of cat tracheal gland cells expressed histological, ultrastructural, and immunological characteristics of mucous cells when cultured on floating gels of rat tail collagen (released-gel cultures) compared with cells cultured on glutaraldehyde-fixed collagen gels (fixed-gel cultures). We therefore collected culture medium from gland cells grown under both culture conditions for determination and comparison of glycoconjugates with characteristics of mucin glycoproteins. Cells were cultured in the presence of [3H]glucosamine, and material of high molecular weight and density (HMD material) was isolated. HMD material from both culture conditions were each resistant to heparitinase and heparinase, whereas 72 and 25% of the radiolabel in released-gel and fixed-gel HMD material, respectively, was resistant to chondroitinase ABC. Material resistant to chondroitinase ABC was analyzed further. Both samples contained a single broad glycoprotein band [relative molecular weight (M(r)) > 250,000] after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and had amino acid profiles similar to airway mucin. The sample from fixed-gel cultures had nearly equal amounts of carbohydrate and protein, was highly enriched in N-acetylglucosamine, contained mannose, displayed little blood group A immunoreactivity, and had few O-linked oligosaccharides. Conversely, the sample from released-gel cultures contained 80% carbohydrate, was composed of monosaccharides characteristic of airway mucins, displayed blood group A immunoreactivity, and contained oligosaccharides O-linked via N-acetylgalactosamine.(ABSTRACT TRUNCATED AT 250 WORDS)
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Park, Yong H., Joshua D. Snook, Iris Zhuang, Guofu Shen, and Benjamin J. Frankfort. "Optimized culture of retinal ganglion cells and amacrine cells from adult mice." PLOS ONE 15, no. 12 (December 7, 2020): e0242426. http://dx.doi.org/10.1371/journal.pone.0242426.

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Cell culture is widely utilized to study the cellular and molecular biology of different neuronal cell populations. Current techniques to study enriched neurons in vitro are primarily limited to embryonic/neonatal animals and induced pluripotent stem cells (iPSCs). Although the use of these cultures is valuable, the accessibility of purified primary adult neuronal cultures would allow for improved assessment of certain neurological diseases and pathways at the cellular level. Using a modified 7-step immunopanning technique to isolate for retinal ganglion cells (RGCs) and amacrine cells (ACs) from adult mouse retinas, we have successfully developed a model of neuronal culture that maintains for at least one week. Isolations of Thy1.2+ cells are enriched for RGCs, with the isolation cell yield being congruent to the theoretical yield of RGCs in a mouse retina. ACs of two different populations (CD15+ and CD57+) can also be isolated. The populations of these three adult neurons in culture are healthy, with neurite outgrowths in some cases greater than 500μm in length. Optimization of culture conditions for RGCs and CD15+ cells revealed that neuronal survival and the likelihood of neurite outgrowth respond inversely to different culture media. Serially diluted concentrations of puromycin decreased cultured adult RGCs in a dose-dependent manner, demonstrating the potential usefulness of these adult neuronal cultures in screening assays. This novel culture system can be used to model in vivo neuronal behaviors. Studies can now be expanded in conjunction with other methodologies to study the neurobiology of function, aging, and diseases.
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Bowers, W. E., and M. R. Berkowitz. "Differentiation of dendritic cells in cultures of rat bone marrow cells." Journal of Experimental Medicine 163, no. 4 (April 1, 1986): 872–83. http://dx.doi.org/10.1084/jem.163.4.872.

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Although dendritic cells (DC) originate from bone marrow, they were not observed in fresh preparations of bone marrow cells (BMC). Likewise, accessory activity was barely measurable in a sensitive assay for this potent function of DC. However, both DC and accessory activity developed when BMC were cultured for 5 d. Based on fractionation before culture, nearly all of the accessory activity could be attributed to only 5% of the total BMC recovered in a low-density (LD) fraction. The LD-DC precursors differed from mature DC in a number of important respects. Removal of Ia+ cells from the LD fraction by panning did not decrease the production of DC when the nonadherent cells were cultured. Thus, the cell from which the DC is derived does not express or minimally expresses Ia antigens, in contrast to the strongly Ia+ DC that is produced in bone marrow cultures. Irradiation of LD cells before culture prevented the development of DC. When irradiation was delayed by daily intervals, progressive increases in the number of DC resulted, up to the fifth day. These findings, together with preliminary autoradiographic data, indicate that cell division has occurred, in contrast to the DC, which does not divide. We conclude that bone marrow-derived DC arise in culture from the division of LD, Ia- precursors.
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Dissertations / Theses on the topic "Cultures Cells"

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Seeley, Marguerite R. "Developmental toxicity of alkylating agents in differentiating cell cultures /." Thesis, Connect to this title online; UW restricted, 1996. http://hdl.handle.net/1773/8464.

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Limoges, Mireille. "Apoptosis in primary cultures of rat Leydig cells." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp02/NQ36586.pdf.

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Dove, N. S. "Electrophysiological studies on primary cultures of skin cells." Thesis, University of Cambridge, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.598614.

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To model sweat gland fluid secretion in vitro, human apocrine glands were isolated from axillary skin and their secretory coil cells were cultured and mounted in an Ussing chamber. Cultured apocrine coil cells exhibited an amiloride-sensitive inward resting short circuit current, suggesting an absorptive phenotype. However, evidence of a chloride-secretory phenotype was also observed: transient inward currents were recorded in response to autonomic secretagogues and inflammatory mediators, and these were not abolished by amiloride, but they were reduced by half in chloride-free buffer. This may indicate a role for chloride secretion in mediating the stimulated currents. Further, sustained inward currents were recorded in 58% of tissues which were attenuated by frusemide, implying the involvement of the Na+/K+/Cl- transporter. Assuming that secondary active chloride transport is responsible for aqueous fluid secretion in the apocrine sweat gland, these data are compatible with some maintenance of an in vivo phenotype in culture. To study the cellular determinants of eccrine sweat gland differentiation, eccrine sweat gland-associated fibroblasts were cultured and compared with derminal fibroblasts from the same subject, demonstrated a different pattern of outgrowth and proliferation. Therefore these cells may represent a novel fibroblast subtype. To improve the differentiation of eccrine sweat gland coil cells in vitro, they were co-cultured with eccrine fibroblasts. Eccrine sweat gland cells grown on a Transwell as a control demonstrated apparent dedifferentiation to a reabsorptive phenotype, indicated by the amiloride-sensitivity of their resting and agonist-stimulated inward short circuit currents. Eccrine cells co-cultured with eccrine fibroblasts, however, demonstrated agonist-stimulated outward currents in the presence of amiloride.
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Montgomery, Sarah Lynn. "Impedance measurement system for embryonic stem cell and embryoid body cultures." Thesis, Atlanta, Ga. : Georgia Institute of Technology, 2008. http://hdl.handle.net/1853/24661.

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Dempsey, Katherine. "Monitoring individual cells within cell cultures using image processing and pattern recognition techniques." Thesis, Keele University, 2017. http://eprints.keele.ac.uk/4179/.

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Cells are the building blocks of the human body which are normally specialised by type in accordance with their function. Human cells interact with each other to form the tissues that make up the body. Consequently, it is important to study the behaviour and interactions of these cells at the microscale level, so that the causes of cellular irregularities can be identified; and, possible treatments can be devised. This project aimed to create algorithms that were capable of tracking a variety of cells types within both single cultures and mixed cultures, and from this generate data that was relevant to current clinical trials. There have been successes in tracking some cells types, most notably articular chondrocytes and spinal disk cells. In terms of data generated there has been successes in a whole variety of different types of clinical trials. The algorithms used here have been able to identify the point of mitosis. They have created a better method of determining neural growth and from this have shown that neurons co-cultured with MCSs can grow in places with neural inhibitors. Through the use of algorithms that can analyse culture in three dimensional structures it has been shown that neurons are more affected by topographical cues than chemical cues in their direction of growth. It has also been shown that vesicles are more likely to appear on smaller back disk cells. In the study of gels, it has been found that the more transparent gels are better for imaging. Finally, it has been shown that MSCs and chondrocytes behave differently when in single and co-cultures. These discoveries would not have been possible without the use of the algorithms that allowed for the study of individual cells within a larger culture.
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Szemiel, Agnieszka M. "Replication of Bunyamwera virus in mosquito cells." Thesis, University of St Andrews, 2011. http://hdl.handle.net/10023/2570.

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The Bunyaviridae family is one of the largest among RNA viruses, comprising more than 350 serologically distinct viruses. The family is classified into five genera, Orthobunyavirus, Hantavirus, Nairovirus, Phlebovirus, and Tospovirus. Orthobunyaviruses, nairoviruses and phleboviruses are maintained in nature by a propagative cycle involving blood-feeding arthropods and susceptible vertebrate hosts. Like most arthropod-borne viruses, bunyavirus replication causes little damage to the vector, whereas infection of the mammalian host may lead to death. This situation is mimicked in the laboratory: in cultured mosquito cells no cytopathology is observed and a persistent infection is established, whereas in cultured mammalian cells orthobunyavirus infection is lytic and leads to cell death. Bunyaviruses encode four common structural proteins: an RNA-dependent RNA polymerase, two glycoproteins (Gc and Gn), and a nucleoprotein N. Some viruses also code for nonstructural proteins called NSm and NSs. The NSs protein of the prototype bunyavirus, Bunyamwera virus, seems to be one of the factors responsible for the different outcomes of infection in mammalian and mosquito cell lines. However, only limited information is available on the growth of bunyaviruses in cultured mosquito cell lines other than Aedes albopictus C6/36 cells. Here, I compared the replication of Bunyamwera virus in two additional Aedes albopictus cell clones, C7-10 and U4.4, and two Aedes aegypti cell clones, Ae and A20, and investigated the impact of virus replication on cell function. In addition, whereas the vertebrate innate immune response to arbovirus infection is well studied, relatively little is known about mosquitoes’ reaction to these infections. I investigated the immune responses of the different mosquito cells to Bunyamwera virus infection, in particular antimicrobial signaling pathways (Toll and IMD) and RNA interference (RNAi). The data obtained in U4.4 cells suggest that NSs plays an important role in the infection of mosquitoes. Moreover infection of U4.4 cells more closely resembles infection in Ae and A20 cells and live Aedes aegypti mosquitoes. My data showed that the investigated cell lines have various properties, and therefore they can be used to study different aspects of mosquito-virus interactions.
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Bell, Cindy Lea. "Transport studies in primary cultures of mouse renal epithelial cells." Thesis, McGill University, 1986. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=75363.

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The Hyp (hypophosphatemic) mouse, a murine homologue of X-linked hypophosphatemia (XLH) in man, is a Mendelian disorder of phosphate (Pi) homeostasis. The mutant genotype is characterized by abnormal Pi transport at the brush border membrane (BBM) of the proximal tubule and a defect in renal metabolism of vitamin D$ sb3$. The exact nature of these defects has not been elucidated.
In order to determine if the defect is intrinsic to the renal cell or dependent upon an extrinsic humoral factor, I established primary cultures of renal epithelial cells from normal and Hyp mouse kidney. The cultures demonstrated several differentiated properties of epithelial cells of the renal proximal tubule, the site of the Pi transport defect in the Hyp mouse.
Primary cultures initiated from Hyp mice had decreased Pi transport (expressed as an uptake ratio, Pi/$ alpha$-MG), and increased production of 24,25 dihydroxyvitamin D$ sb3$. These results provide evidence for the intrinsic nature of the primary defect in the Hyp mouse.
This appears to be the first time that expression of a mutant transport gene has been demonstrated in cultured renal cells.
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Zhang, Fan. "Electric and electrochemical responses of adherent cells : application of microfabrication technologies." Paris 6, 2011. http://www.theses.fr/2011PA066194.

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La connaissance des comportements cellulaires tels que l'adhésion, migration et prolifération cellulaire est importante pour l'ingénierie tissulaire et de l'implantologie. Ce travail de thèse a été développé pour obtenir une vision plus claire sur les activités cellulaires in-vitro en utilisant des substrats micro/nanofabriqués et des méthodes d’analyses électriques/électrochimiques. Des substrats avec des motifs divers et variés, incluant des nanoélectrodes de haute densité et des microstructures à trois dimensions, ont été obtenu pour culture cellulaire et analyse par méthodes électriques ou électrochimiques. L’intégration de ces structures dans un dispositif microfluidique a été également démontrée. L'adhésion, migration et prolifération cellulaire ainsi que l'activité métabolique des cellules ont été étudié par mesure de voltamétrie cyclique et d'impédance électrique. En combinaison avec les techniques optiques pour l'observation de la morphologie cellulaire et la densité de cellules, les mesures électriques ou électrochimiques nous ont permis d’étudier des nouveaux effets de substrats ou électrodes micro et nano-structurés sur la culture cellulaire et les activités métaboliques de cellules en culture
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Pucihar, Gorazd. "Induced transmembrane voltage and electropermeabilization of cells in cultures in vitro." Toulouse 3, 2006. http://thesesups.ups-tlse.fr/1038/.

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Quand une cellule est exposée à un champ électrique externe, la tension électrique transmembranaire (ITV) est modifiée. Pendant l'exposition, l' ITV se superimpose au potentiel de repos (RTV) et quand la somme des deux tensions excède une valeur critique, la perméabilité de la membrane cellulaire augmente transitoirement localement. Ce phénomène est désigné comme electropermeabilisation. Dans beaucoup d'applications de l'electropermeabilisation une permeabilisation efficace et en même temps réversible est essentielle. Ainsi, une prédiction de l'expérience, qui implique l'évaluation de l'amplitude de l'ITV pour déclencher la permeabilisation, est exigée. Le problème est critique dans des tissus, où la géométrie cellulaire est plus compliquée, les cellules sont assez proches pour affecter le champ électrique autour d'elles et elles sont souvent connectées entre elles. Dans tous ces cas, une description analytique de l'ITV n'est en général pas accessible et des méthodes numériques sont ainsi souvent la seule approche envisageable. En raison de la complexité de la structure tissulaire, les modèles sont macroscopiques, et on ne considère pas la structure cellulaire détaillée, ou en cas des modèles microscopiques, les modèles sont construits utilisant des formes géométriques simples (des semi-sphères, des cubes). Pour mieux comprendre comment le champ électrique interagit avec des tissus, nous avons construit les modèles microscopiques réalistes de cellules irrégulièrement formées, des groupes de telles cellules et des suspensions denses. Le travail a alors été développé sur le plan expérimental au niveau de la cellule isolée. Les mesures de cinétique de transport de membrane ont montré que l'electropermeabilisation avec des amplitudes d'impulsion ou des durées d'impulsion progressivement croissantes conduit à des transports accrus dans des cellules. Une large augmentation a été observée dans les milisecondes après le début d'une impulsion, suivie par une augmentation de fluorescence progressive. Les résultats mesurés sur un intervalle de temps de 400 µ S ont révélé que le transport à travers la membrane permeabilisée ne peut être détecté que 100 µ S après le début de l'impulsion. En plus, une dynamique différente d'augmentation de fluorescence pendant et après l'impulsion a été observée
When a biological cell is exposed to an external electric field, induced transmembrane voltage (ITV) forms on its membrane. During the exposure, ITV superimposes to the native or resting transmembrane voltage (RTV) and when the sum of both voltages exceeds some threshold value, the permeability of the cell membrane in these regions transiently increases. This phenomenon is termed electropermeabilization. In many applications of electropermeabilization an efficient and at the same time reversible permeabilization is essential (e. G. DNA electrotransfer). Thus, a careful planning of the experiment, which involves the estimation of the amplitude of ITV leading to cell permeabilization, is required. The problem arises in case of tissues, where cell geometry is more complicated, cells are close enough to affect the electric field around each other, and they are often connected with pathways between them. In all these cases, an analytical description of ITV is in general not attainable and numerical methods are often the only feasible approach. Due to the complexity of tissue structure, numerical models are either macroscopic, where detailed cell structure is notconsidered, or in case of microscopic models, the models are constructed using simple geometrical shapes (semi-spheres, cubes). To better understand how the electric field interacts with tissues on a microscopic (single cell) level, which in turn determines the macroscopic behavior of the tissue, we constructed realistic microscopic models of irregularly shaped cells, clusters of such cells, and dense suspensions. Regarding the shape, density and connections between cells, these cell assemblies are in their complexity close to tissues. First, the amplitude of resting transmembrane voltage of cells used in the study was determined. Next, calculations of ITV were performed on models of single spherical, single attached cells, and cell clusters and they were compared to measurements of ITV on the same cells, from which the models were constructed. The course of electropermeabilization of these cells was then monitored and the results were compared with measurements and calculations of ITV. In a separate experiment, a detailed investigation of kinetics of molecular transport into cells after permeabilization was performed. Similarly, for dense cell suspensions, the ITV calculated on a model of suspension was compared with the fraction of permeabilized cells measured in suspensions with increasing cell densities. Measurements of resting transmembrane voltage (RTV) were performed by means of a slow potentiometric fluorescent dye TMRM on different cell lines in culture media and media with progressively decreasing conductivities. ITV was measured on single spherical cells, single irregularly shaped cells, and cell clusters with a fast potentiometric fluorescent dye di-8- ANEPPS. The cross-section fluorescence images of the same cells on which the measurements of ITV were performed, were used to construct realistic numerical models of cells and the ITV on these models was then calculated with finite elements method. Finitethickness, nonzero conductivity cell membrane in the model was replaced by a boundary condition in which a specific surface conductivity was assigned to the interface between the cell interior and the exterior. Electropermeabilization of cells was followed by monitoring thechanges in intracellular fluorescence of membrane-impermeant fluorescent dye Propidium Iodide. Measurements of RTV showed that in physiological conditions (cells in culture medium) and in the presence of pulsing buffer, RTV on investigated cell lines is low (between -4 and -35 mV for suspended cells and between -18 and -27 mV for attached cells). Therefore, in experiments involving electropermeabilization ITV can be used as a rough approximate of the total voltage on the membrane, while RTV can be neglected. RTV in cells in media with decreasing conductivities gradually decreased, but less than expected from theoretical calculations. This was partly attributed to overestimated intracellular concentration of potassium. However, it is also possible that the method for measuring RTV, although reported as efficient, was not suitable for these experiments. Measurements of ITV on single spherical cells, single attached cells, and cell clusters were in qualitative agreement with results of numerical calculations, while in some cases discrepancies in measured and calculated amplitudes could be observed. This was attributed to variations of the slope of calibration curve, the differences between the actual and implemented parameters of the model, physiological state of cells, and experimental setup. In addition, we observed that at pulse parameters used in measurements of ITV, cells in clusters behaved as electrically connected, i. E. A cluster acted as one giant cell. Numerical calculations on models of cells where cell membrane was replaced with a boundary condition resulted in considerably lower number of mesh elements and consequently shorter time needed to solve the problem. We also demonstrated that calculations of ITV on simplified models of irregularly shaped cells can lead to considerable deviations from ITV calculated on a realistic model. Electric field orientation affects the amplitude and distribution of calculated ITV and consequently permeabilization. Namely, cells oriented with their longer axis parallel to the field are more likely to get permeabilized than the same cells oriented perpendicularly to the field. Comparison of measured and calculated ITVs with observations of electropermeabilization on single spherical and single attached cells confirmed that permeabilization occurs in those regions of the membrane, where the absolute value of ITV is the highest (the regions facing the electrodes). Additional experiments performed on single spherical cells showed that during and immediately after the pulse, the fluorescence from cells increases asymmetrically if unipolar pulses were delivered, while symmetrical fluorescence was observed for bipolar pulses. These observations were attributed to electrophoretical effect of the pulse. On a longer time scale, asymmetry in fluorescence was still observed, even for bipolar pulses, and we did not find any reasonable explanation for that. Critical value of ITV, at which permeabilization occurs, was calculated from the polar angle of permeabilization measured immediately after the pulse and was found to be approximately 450 mV, in agreement with reported critical thresholds. Permeabilization results obtained on cell clusters showed that cells in clusters, atpulse parameters used in these experiments, behaved as electrically insulated and were permeabilized individually. This is in contradiction to what we observed during measurements of ITV (i. E. With longer, low voltage pulses), where cells in clusters behaved as electrically connected, and was assumed to be the result of opening and closing of gap junctions at different pulse parameters. Measurements of kinetics of membrane transport showed that electropermeabilization with progressively increasing pulse amplitudes or pulse durations results in increased dye transport into cells. A sharp increase was observed miliseconds after the onset of a pulse, followed by a moderate additional fluorescence increase. Results measured on a time interval of 400 µs revealed that the transport across the permeabilized membrane can be detected within 100 µs after the onset of the pulse. Besides, different dynamics of fluorescence increase was observed during and immediately after the pulse. Experiments carried out on dense cell suspensions showed that with increasing cell density (from 10×106 cells/ml to 400×106 cells/ml) the fraction of permeabilized cells decreased by approximately 50%. We attributed this to the changes in the local electric field, which lead to a decrease in the amplitude of ITV. The uptake of Propidium Iodide also decreased with cell density, but by a larger amount than expected from permeabilization results. We supposed that the additional decrease in fluorescence was mainly due to cell swelling after permeabilization, which reduced extracellular dye availability to the permeabilized membrane and hindered the dye diffusion into the cells. Resealing of cells appeared to be slower in dense suspensions, which can also be attributed to cell swelling resulting from electropermeabilization
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YOSHITO, DANIELE. "Cultivo e irradiação de fibroblastos humanos em meio enriquecido com lisado de plaquetas para obtenção de camada de sustentação em cultura de células da epiderme." reponame:Repositório Institucional do IPEN, 2011. http://repositorio.ipen.br:8080/xmlui/handle/123456789/9959.

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Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP
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Books on the topic "Cultures Cells"

1

Freiburg Focus on Biomeasurement (8th 1993 Freiburg, Germany). Methods and applications of single cells and cell cultures in physiology and pharmacology: 8th Freiburg Focus on Biomeasurement, February 15th and 16th, 1993. Buchenbach: Biomesstechnik-Verlag, 1994.

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Chaddah, Maya Rani. Characterization of srem cells from long-term bone marrow cultures. Ottawa: National Library of Canada, 1993.

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Biology of normal proliferating cells in vitro: Relevance for in vivo aging. Basel: Karger, 1988.

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Ryan, Martyn J. The effect of hydrodynamic stress on plant cell cultures in turbulent jet flows. Dublin: University College Dublin, 1997.

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Bone research protocols. 2nd ed. New York: Humana Press, 2012.

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S, Ambesi-Impiombato F., and Perrild H, eds. FRTL-5 today: Proceedings of the First International Workshop on Characterization and Standardization of an In Vitro Thyroid Cell System, Udine, Italy, 26-28 October 1988. Amsterdam: Excerpta Medica, 1989.

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Ozolinš, Terence Robert Stanislavs. Interspecies co-culture of embryos and maternal hepatocytes: An in vitro model of phenytoin embryotoxicity. Toronto, Ont: Faculty of Pharmacy, University of Toronto, 1990.

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Edwards, M. J. ATCC microbes & cells at work: An index to ATCC strains with special applications. Rockville, Md: American Type Culture Collection, 1988.

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Plant cell culture protocols. 3rd ed. New York: Springer, 2012.

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Koller, Manfred R. Human Cell Culture: Volume IV: Primary Hematopoietic Cells. Dordrecht: Kluwer Academic Publishers, 2002.

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Book chapters on the topic "Cultures Cells"

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Healy, Lyn, and Ludmila Ruban. "Culture Adaptation and Abnormal Cultures." In Atlas of Human Pluripotent Stem Cells in Culture, 167–75. Boston, MA: Springer US, 2014. http://dx.doi.org/10.1007/978-1-4899-7507-2_10.

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Choi, Jeong-Woo, Gyu Heon Cho, Sang Yo Byun, and Dong-Il Kim. "Integrated Bioprocessing for Plant Cell Cultures." In Plant Cells, 63–102. Berlin, Heidelberg: Springer Berlin Heidelberg, 2001. http://dx.doi.org/10.1007/3-540-45302-4_3.

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Miranda, Joana, Manuel J. T. Carrondo, and Paula M. Alves. "3D Cultures: Effect on the Hepatocytes Functionality." In Cells and Culture, 171–76. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-90-481-3419-9_28.

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Sagmeister, Patrick, Mohammadhadi Jazini, Joachim Klein, and Christoph Herwig. "Bacterial Suspension Cultures." In Industrial Scale Suspension Culture of Living Cells, 40–93. Weinheim, Germany: Wiley-VCH Verlag GmbH & Co. KGaA, 2014. http://dx.doi.org/10.1002/9783527683321.ch01.

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Spahr, R., and H. M. Piper. "Microcarrier Cultures of Endothelial Cells." In Cell Culture Techniques in Heart and Vessel Research, 220–29. Berlin, Heidelberg: Springer Berlin Heidelberg, 1990. http://dx.doi.org/10.1007/978-3-642-75262-9_15.

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Mukherjee, Nabanita, Karoline A. Lambert, David A. Norris, and Yiqun G. Shellman. "Enrichment of Melanoma Stem-Like Cells via Sphere Assays." In Methods in Molecular Biology, 185–99. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1205-7_14.

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AbstractSphere assays are widely used in vitro techniques to enrich and evaluate the stem-like cell behavior of both normal and cancer cells. Utilizing three-dimensional in vitro sphere culture conditions provide a better representation of tumor growth in vivo than the more common monolayer cultures. We describe how to perform primary and secondary sphere assays, used for the enrichment and self-renewability studies of melanoma/melanocyte stem-like cells. Spheres are generated by growing melanoma cells at low density in nonadherent conditions with stem cell media. We provide protocols for preparing inexpensive and versatile polyHEMA-coated plates, setting up primary and secondary sphere assays in almost any tissue culture format and quantification methods using standard inverted microscopy. Our protocol is easily adaptable to laboratories with basic cell culture capabilities, without the need for expensive fluidic instruments.
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F�rster, Eckart, Marlene Bartos, and Shanting Zhao. "Hippocampal Slice Cultures." In New Methods for Culturing Cells from Nervous Tissues, 1–11. Basel: KARGER, 2005. http://dx.doi.org/10.1159/000083427.

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Kapfhammer, Josef P. "Cerebellar Slice Cultures." In New Methods for Culturing Cells from Nervous Tissues, 74–81. Basel: KARGER, 2005. http://dx.doi.org/10.1159/000083443.

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Linden, J. C., J. R. Haigh, N. Mirjalili, and M. Phisaphalong. "Gas Concentration Effects on Secondary Metabolite Production by Plant Cell Cultures." In Plant Cells, 27–62. Berlin, Heidelberg: Springer Berlin Heidelberg, 2001. http://dx.doi.org/10.1007/3-540-45302-4_2.

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Gritti, Angela, Rossella Galli, and Angelo L. Vescovi. "Clonal Analyses and Cryopreservation of Neural Stem Cell Cultures." In Neural Stem Cells, 173–84. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-59745-133-8_14.

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Conference papers on the topic "Cultures Cells"

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Berthier, R., A. Duperray, O. Valiron, M. Prenant, I. Newton, and A. Schweitzer. "MEGAKARYOCYTIC DEVELOPMENT IN LIQUID CULTURES OF CRYOPRESERVED LEUKOCYTE STEM CELL CONCENTRATES FROM CHRONIC MYELOGENOUS LEUKEMIA PATIENTS." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644622.

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The proliferation and differentiation of human megakaryocytes in liquid culture has been obtained using cryopreserved light density blood cell concentrates from chronic myelogenous leukemia (CML) patients. These cryopreserved leukocytes concentrates contain a large number of viable granulo-monocytic, erythroid and megakaryocytic committed stem cells. A high number of spontaneous megakaryocytic colonies was observed in semisolid cultures plated with the CML leukocytes concentrates. A liquid culture system using RPMI 1640 supplemented with 20% human plasma (HP) has been defined where maturing megakaryocytes make up 20 to 60% of the total cells after 14 days of incubation. The same cell suspension cultured in medium supplemented with 20% foetal calf serum (FCS) showed poor megakaryocytic cell development. The megakaryocytic nature of the cells produced in HP supplemented cultures was confirmed by cytological studies and indirect immunofluorescence labeling using monoclonal antibodies (MoAb) against membrane platelet GPIb and Ilbllla, and intracellular antigens like fibrinogen and von Willebrand factor.Ploidy of the cultured cells was studied after labeling with propidium iodide and the DNA fluorescence determined using the fluorescence activated cell sorter (FACSIV). Peaks of 8N, 16N and 32N cells were observed from HP supplemented cultures representing about 20% of the cells reacting with a GP11b111 a MoAb, while very few cells greater than 4N were observed in FCS supplemented cultures. The megakaryocytes produced in HP cultures could be further enriched by cell sorting on the FACSIV after labeling with an anti-IIbIIIa MoAb. Depending on the initial megakaryocytic concentration of the cells cultured, one to 2 é 106 megakaryocytes per hour could be harvested. Thus, cryopreserved CML blood stem cell concentrates seem to offer a reproducible source of human megakaryocytes which retain their capacity to proliferate and differentiate in liquid cultures supplemented with human plasma. These megakaryocytes can be used for the study of platelet glycoprotein biosynthesis as well as the regulation of megakaryocytopoiesis.
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Isu, Giuseppe, Diana Massai, Giulia Cerino, Diego Gallo, Cristina Bignardi, Alberto Audenino, and Umberto Morbiducci. "A Novel Perfusion Bioreactor for 3D Cell Culture in Microgravity Conditions." In ASME 2013 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/sbc2013-14502.

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Cell suspension culture methods based on the generation of microgravity environment are widely used in regenerative medicine for (1) the production of native-like three-dimensional (3D) cell aggregates and engineered tissues [1,2,3], for (2) low cost scalable cell expansion and long-term cell viability maintenance [4,5], and for (3) guiding differentiation of stem cells (SCs) [6]. The generation of a microgravity environment for 3D cell cultures, mimicking the native environment, promotes spatial freedom, cell growth, cell-cell interaction and improves mass transfer and cell exposure to nutrients. Nowadays, microgravity cell cultures are obtained by using stirred or rotating bioreactors, but both devices suffer from limitations: stirring bioreactors generate non-physiological shear stresses, which could damage cultured cells, interfere with SC pluripotency, and limit reproducibility of the culture process; rotating bioreactors are expensive devices due to the complex technological solutions adopted for obtaining rotation [5].
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Singh, Ankur, Shalu Suri, Ted T. Lee, Jamie M. Chilton, Steve L. Stice, Hang Lu, Todd C. McDevitt, and Andrés J. Garcia. "Adhesive Signature-Based, Label-Free Isolation of Human Pluripotent Stem Cells." In ASME 2012 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2012. http://dx.doi.org/10.1115/sbc2012-80044.

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Generation of human induced pluripotent stem cells (hiPSCs) from fibroblasts and other somatic cells represents a highly promising strategy to produce auto- and allo-genic cell sources for therapeutic approaches as well as novel models of human development and disease1. Reprogramming protocols involve transduction of the Yamanaka factors Oct3/4, Sox2, Klf4, and c-Myc into the parental somatic cells, followed by culturing the transduced cells on mouse embryonic fibroblast (MEF) or human fibroblast feeder layers, and subsequent mechanical dissociation of pluripotent cell-like colonies for propagation on feeder layers1, 2. The presence of residual parental and feeder-layer cells introduces experimental variability, pathogenic contamination, and promotes immunogenicity3. Similar to human embryonic stem cells (hESCs), reprogrammed hiPSCs suffer from the unavoidable problem of spontaneous differentiation due to sub-optimal feeder cultures4, growth factors5, and the feeder-free substrate6. Spontaneously differentiated (SD)-hiPSCs display reduced pluripotency and often contaminate hiPSC cultures, resulting in overgrowth of cultures and compromising the quality of residual pluripotent stem cells5. Therefore, the ability to rapidly and efficiently isolate undifferentiated hiPSCs from the parental somatic cells, feeder-layer cells, and spontaneously differentiated cells is a crucial step that remains a bottleneck in all human pluripotent stem cell research.
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Hunter, N. R., I. R. MacGregor, J. Dawes, and D. S. Pepper. "MICROCARRIER CULTURE OF HUMAN ENDOTHELIAL CELL TYPES - A SOURCE OF METABOLITES." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643348.

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The production of human endothelial cell secretory products in amounts sufficient for biochemical studies is largely restricted by the culture growth area. Conventional flat bed systems yield at best 20-30 x 106 cells per 180cm2 culture flask. To overcome this problem, cells may be grown on Cytodex 3 microcarriers allowing large numbers of cells to be grown and conditioned in small culture volumes. A typical microcarrier unit will contain 200-300 x 106 cells and may be expanded in excess of 1000 x 106 cells at confluence. High viability (95%) and recovery (70-80%) in sub-culturing of microcarrier to microcarrier culture can be achieved with careful management of culture conditions and brief exposure to enzymes.Human umbilical artery and vein, and saphenous vein endothelial cells were prepared and grogn on microcarrier cultures to cell populations of 200-450 x 106 cells and conditioned for 14 day periods in serum-free media.The production profiles of several endothelial cell proteins including thrombospondin (TSP), von Willebrand Factor (vWF) and issue plasminogen activator (t-PA) were measured by radioimmunoassay under these conditions, and demonstrate the use of microcarrier cultures in producing milligram quantities of engothelial cell protein. For example, a HUVEC culture of 200 x 106 cells conditioned with serum-free media for 14 days yielded a total of 6.9mg TSP, 0.7mg vWF and 48.9ug t-PA. In this laboratory one such application of the system was the purification of endothelial proteins in amounts sufficient for immunisation of mice prior to the production of monoclonal antibodies and for subsequent characterisation.
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Mignone, Lindsay F., Shirley Masand, Jeffrey D. Zahn, and David I. Shreiber. "A Simple, Cost-Effective Method to Improve Cell Viability in Microniche Culture Systems." In ASME 2010 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2010. http://dx.doi.org/10.1115/sbc2010-19189.

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Microfluidic networks are increasingly used to generate custom microenvironmental niches for cell culture and assays of cellular behavior. Perfusion systems are typically required to overcome diffusive limitations associated with culturing cells longer than a few hours when nutrient delivery, oxygen delivery and metabolic waste removal are required to maintain cell viability. In addition to the added complexity of experimental methods, perfusion systems can result in nonuniform nutrient delivery and subject cells to shear stresses, which may alter cell behavior and possibly cause cell death. In particular, when culturing cells within hydrogel scaffold-filled networks, as may be done in micro-tissue engineering, the need for perfusion culture also increases the likelihood of a destructive bubble entering the network. Moreover, analysis of micro-cultures frequently entails labelling with antibodies and/or fluorescent probes, which again requires controlled perfusion of the various reagents through the network. We have developed a simple technique to preserve cell viability and simplify labeling within microscale cultures without the need for perfusion. Instead of bonding a microfluidic network to glass, PDMS, or another impermeable substrate, the network is bonded to a semi-permeable microdialysis membrane, which allows free exchange of oxygen, proteins, nutrients, and waste between the microfluidic channels and culture media in static culture plates.
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Korner, G., and Thorir D. Bjornsson. "INTRACELLULAR REGULATION OF TRANSGLUTAMINASE IN INTACT AND H202 INJURED CLONED BOVINE ENDOTHELIAL CELLS." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1642863.

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Vascular endothelial cells are normally in a quiescent, nonproliferating state. After intimal injury, they are likely to undergo proliferation. We found that cloned bovine aortic endothelial cells (EC) in culture contain high activity of tissue-type transglutaminase (TG). The enzyme was Ca++ dependent, the Km and vmax for putrescine were 0.203 and 18.5 nmol/min/mg protein. Primary amines were capable of inhibiting its activity but not methylated dansylcadaverine. EC and TG molecular weight estimated by gel filtration or by SDS-PAGE, was 88±5,000 Kd. Immunologically it was cross-reactive with purified rat liver TG. TG activity and antigen were found to increase significantly, when the cultures reached confluence and even more when cultures arrested at G0/G1 state by serum depletion. The half-life of EC-TG in G0/G1 arrested cultures was 128 minutes, determined in the presence cycloheximide; no decrease was observed in confluent cultures, indicating active accumulation of TG at the non-pro-liferative state. Most of cellular TG activity was found in the 15,000 g soluble fraction, with the crude membrane fraction containing 4-22% of the total TG activity, depending on the state of cell proliferation. However, immunoblots revealed that the membrane fraction contained similar amounts of TG antigen as did the soluble fraction. When the membranes were treated with detergents (0.1% SDS, 0.5 M NaCl, 75 mM KSCN, 75 mM dithio-threitol), the activity increased significantly, resulting in as much as 4- to 7-fold increases above control. Similar treatment of the soluble fraction did not increase TG activity. This data strongly suggests that the cell membranes contain a latent or cryptic form of TG that can undergo activation. Indeed, when cultures were injured by exposing them to H2O2 (10-10 mM) , a rapid 75-200% increase in TG activity was observed. One hour later, the activity was at control level. If protein synthesis inhibitors were present, no decrease was noted up to 8 hours. It is proposed that significant amounts of TG is stored in an inactive form in endothelial cell membranes, and that under different conditions, e.g., cell injury, activation and translocation of the enzyme can occur, out of and back into the cells membrane.
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7

Schaub, R. G., C. J. Dunn, D. E. Tracey, W. E. Fleming, and M. D. Burdick. "THROMBOTIC AND INFLAMMATORY CHANGES IN ENDOTHELIAL CELLS INCUBATED WITH LEUKOCYTES." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1642860.

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Adhesion of leukocytes (WBC's) to vascular endothelial cells (EC's) is a component of inflammation, thrombosis and atherosclerosis. The purpose of this study was to assess the effect of WBC adhesion on the EC contribution to these pathologic events. Human WBC's were isolated and co-incubated with cultured human umbilical cord EC's. Supernatants and cell lysates (4 wells × 2) were obtained at 0.5,1,2, and 4 hours of incubation. EC's and WBC's (5 × 105) were-incubated alone or in combination. Supernatants and cell lysates were assayed for leukotriene B4 (LTB4), the thromboxane metabolite thromboxane B2 (TXB2) and the prostacyclin metabolite 6-keto prostaglandin FT alpha (6 keto) by RIA. Cell lysates were analyzed for cell associated procoagulant activity (PCA) by an APTT procedure, for plasminogen activator inhibitor (PAI) by an amidolytic assay and for IL-1 by a T-cell co-stimulator assay. Cellular and supernatant LTB4 was unmeasurable for both WBC's and WBC/EC cultures. WBC TXB2 showed a time dependent elevation which was unaffected by EC's. IL-1 activity was measurable at 2 hours and reach 14 U/ml in WBC's and 6 U/ml in EC/WBC cultures. Co-incubation of WBC's with EC's induced a 200% increase in both supernatant and cell associated 6 keto concentrations compared to EC's incubated alone. EC's and WBC's produced no PCA when incubated alone. PCA activity of the EC/WBC co-cultures was measurable at 2 hours and was 300-1500 U/ml after 4 hours. Coincubated EC's had a 50% decrease in cell PAI, suggesting an increased release of inhibitor from the cells. The prostacyclin and PAI release -along with the delayed expression of PCA activity are responses similar to those expected after EC exposure to cytokines. A source of these cytokines appears to be the WBC's which secreted measurable amounts of IL-1. WBC released IL-1 was sufficient to induce biochemical changes in EC's which can stimulate coagulation (PCA synthesis), inhibit fibrinolysis (PAI release), and enhance inflammation (prostacyclin synthesis). These results suggest that the release of WBC IL-1 can be sufficient to produce pro-thrombotic and inflammatory changes in EC's which are similar to those observed with the addition of exogenous IL-1 to EC cultures.
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Vokac, KA, J. Ferrars, and RR Montgomery. "RISTOCETIN-INDUCED ENDOTHELIAL CELL BINDING OF PLASMA VON WILLEBRAND FACTOR." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1642913.

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Our laboratory previously used a technique of labeling plasma von Willebrand factor (vWf) with radiolabeled AVW1 - a “ neutral” monoclonal antibody to vWf. This technique has been used to study the binding of plasma vWf to platelets in the plasma milieu. Studies by several laboratories including ours have demonstrated structural glycoproteins on endothelial cells that are analogous to platelet GP Ilb/IIIa and we have shown that the platelet alloantigen Pl-Al is expressed on the surface of cultured endothelial cells. We undertook this study to evaluate the binding of plasma vWf to cultured endothelial cells in confluent monolayer cultures using the “ neutral” monoclonal antibody technique. Plasma vWf was “ labeled” using trace quantities of radiolabeled AVW-1. We then added 80,000 cpm of monoclonal-labeled plasma to 48 well culture plates containing confluent secondary cultures of human umbilical vein endothelial cells. Following the addition of ristocetin, the plates were incubated for 1 hour at room temperature, centrifuged, and the count8 bound and the counts remaining in the supernate were determined. In the presence of ristocetin, 67.5% of the labeled vWf bound to the endothelial cells. When “ labeled“ severe von Willebrand plasma was used or when ristocetin was omitted, less than 5% of the counts bound. Controls using mouse serum or excess mouse IgG to rule out Fc receptor binding and controls to evaluate binding to the subcellular matrix were performed and demonstrated this binding to be vWf and cell surface dependent. Unlike platelet vWf binding, this binding was not inhibited by monoclonal or polyclonal antibodies to platelet GPIb. We studied plasma from patients with type I, a variant of type I, and type Ila vWd and found normal binding with the type I plasma, but reduced binding with the type I variant plasma (14.5%) and the type Ila plasma (7.1%). AVW3, a monoclonal antibody to vWf that blocks vWf binding to platelet GPIb, blocked vWf binding to endothelial cells. Endothelial cells, like platelets, have the ability to bind plasma von Willebrand factor in the presence of ristocetin. This phenomenon occurs on the surface of endothelial cells in culture. Qualitative and quantitative reductions of this vWf binding are found with the plasma of patients with von Willebrand’s disease.
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Kim, Minwook, Jason A. Burdick, and Robert L. Mauck. "Influence of Chondrocyte Zone on Co-Cultures With Mesenchymal Stem Cells in HA Hydrogels for Cartilage Tissue Engineering." In ASME 2012 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2012. http://dx.doi.org/10.1115/sbc2012-80859.

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Mesenchymal stem cells (MSCs) are an attractive cell type for cartilage tissue engineering in that they can undergo chondrogenesis in a variety of 3D contexts [1]. Focused efforts in MSC-based cartilage tissue engineering have recently culminated in the formation of biologic materials possessing biochemical and functional mechanical properties that match that of the native tissue [2]. These approaches generally involve the continuous or intermittent application of pro-chondrogenic growth factors during in vitro culture. For example, in one recent study, we showed robust construct maturation in MSC-seeded hyaluronic acid (HA) hydrogels transiently exposed to high levels of TGF-β3 [3]. Despite the promise of this approach, MSCs are a multipotent cell type and retain a predilection towards hypertrophic phenotypic conversion (i.e., bone formation) when removed from a pro-chondrogenic environment (e.g., in vivo implantation). Indeed, even in a chondrogenic environment, many MSC-based cultures express pre-hypertrophic markers, including type X collagen, MMP13, and alkaline phosphatase [4]. To address this issue, recent studies have investigated co-culture of human articular chondrocytes and MSCs in both pellet and hydrogel environments. Chondrocytes appear to enhance the initial efficiency of MSC chondrogenic conversion, as well as limit hypertrophic changes in some instances (potentially via secretion of PTHrP and/or other factors) [5–7]. While these findings are intriguing, articular cartilage has a unique depth-dependent morphology including zonal differences in chondrocyte identity. Ng et al. showed that zonal chondrocytes seeded in a bi-layered agarose hydrogel construct can recreate depth-dependent cellular and mechanical heterogeneity, suggesting that these identities are retained with transfer to 3D culture systems [8]. Further, Cheng et al. showed that differences in matrix accumulation and hypertrophy in zonal chondrocytes was controlled by bone morphogenic protein [9]. To determine whether differences in zonal chondrocyte identity influences MSC fate decisions, we evaluated functional properties and phenotypic stability in photocrosslinked hyaluronic acid (HA) hydrogels using distinct, zonal chondrocyte cell fractions co-cultured with bone marrow derived MSCs.
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10

Wynne, Rosalind, and Sabrina Ahmed. "Fabrication Considerations for Bridged Microfluidic Cell Cultures." In ASME 2018 Conference on Smart Materials, Adaptive Structures and Intelligent Systems. American Society of Mechanical Engineers, 2018. http://dx.doi.org/10.1115/smasis2018-7983.

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A novel bridged-microfluidic for cell-based assays was developed by combining a microstructured optical fiber (MOF) with a microfluidic network with the purpose of continuously monitoring the state of hepatocellular carcinoma (HepG2) cells. In this configuration a solid core MOF with channels in the cladding serves as a bridge for cell transport as well as an evanescent wave-based monitoring system to detect cells labeled with fluorescent nanomaterials. The device was fabricated by positioning an MOF to bridge two polydimethylsiloxane (PDMS) microfluidic networks. Alignment strategies and pressurization considerations to produce this system are presented. Pump systems that support fluid transport through the MOF demonstrated the tendency of flow rate fluctuations even for constant microfluidic pump rates. Spectroscopic measurements confirm the delivery and motion of cells between the two neighboring microfluidic chips. The linewidth of the spectra demonstrated oscillations that were consistent with pressure broadening caused by hydrodynamic fluctuations. Fluctuations in the microfluidic flow ranging from 0.005 to 0.016 Hz were observed. These results are consistent with theoretical principles and provide important information regarding syringe pump artifacts, i.e. fluctuations, observed during spectroscopic measurements in MOF/microfluidic systems.
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Reports on the topic "Cultures Cells"

1

Kahler, David W., and Carmen M. Arroyo. Normal Human Astrocyte Instructions for Initiation of Cultures from Cryopreserved Cells and Subculture. Fort Belvoir, VA: Defense Technical Information Center, October 2004. http://dx.doi.org/10.21236/ada442897.

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2

Peehl, Donna M. Development of Methodology to Maintain Primary Cultures of Normal and Malignant Human Prostatic Epithelial Cells In Vivo. Fort Belvoir, VA: Defense Technical Information Center, February 2008. http://dx.doi.org/10.21236/ada484336.

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3

Malik, Abir, D. Lam, H. A. Enright, S. K. G. Peters, B. Petkus, and N. O. Fischer. Characterizing the Phenotypes of Brain Cells in a 3D Hydrogel Cell Culture Model. Office of Scientific and Technical Information (OSTI), August 2018. http://dx.doi.org/10.2172/1466140.

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4

Novaro, Virginia, and Mina BIssell. A Cell Culture Model for Understanding Estrogen Receptor Regulation in Normal and Malignant Cells. Fort Belvoir, VA: Defense Technical Information Center, October 1998. http://dx.doi.org/10.21236/ada373393.

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5

Grego, Sonia, Edward R. Dougherty, Francis J. Alexander, Scott S. Auerbach, Brian R. Berridge, Michael L. Bittner, Warren Casey, et al. Systems Biology for Organotypic Cell Cultures. Office of Scientific and Technical Information (OSTI), August 2016. http://dx.doi.org/10.2172/1313549.

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6

Merkle, Carrie J. Studies on Breast Cancer Cell Interactions with Aged Endothelial Cells in Culture and Rat Models. Fort Belvoir, VA: Defense Technical Information Center, May 2006. http://dx.doi.org/10.21236/ada455981.

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7

Richmond, Robert C. Cell and Molecular Biology of Ataxia Telangiectasia Heterozygous Human Mammary Epithelial Cells Irradiated in Culture. Fort Belvoir, VA: Defense Technical Information Center, September 2002. http://dx.doi.org/10.21236/ada412826.

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8

Eisinger, Magdalena. Wound Healing by Cultured Skin Cells and Growth Factors. Fort Belvoir, VA: Defense Technical Information Center, June 1994. http://dx.doi.org/10.21236/ada284593.

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9

Karin, M. The molecular basis for uv response of cultured human cells. Office of Scientific and Technical Information (OSTI), December 1991. http://dx.doi.org/10.2172/10104960.

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10

Karin, M. The molecular basis for uv response of cultured human cells. Office of Scientific and Technical Information (OSTI), January 1991. http://dx.doi.org/10.2172/6261995.

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