Academic literature on the topic 'Angiogenic cells'

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Journal articles on the topic "Angiogenic cells"

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Sadagopan, Sathish, Neelam Sharma-Walia, Mohanan Valiya Veettil, Virginie Bottero, Rita Levine, Richard J. Vart, and Bala Chandran. "Kaposi's Sarcoma-Associated Herpesvirus Upregulates Angiogenin during Infection of Human Dermal Microvascular Endothelial Cells, Which Induces 45S rRNA Synthesis, Antiapoptosis, Cell Proliferation, Migration, and Angiogenesis." Journal of Virology 83, no. 7 (January 21, 2009): 3342–64. http://dx.doi.org/10.1128/jvi.02052-08.

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ABSTRACT Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) is associated with the angioproliferative KS lesions characterized by spindle-shaped endothelial cells, inflammatory cells, cytokines, growth factors, and angiogenic factors. De novo KSHV infection of human microvascular dermal endothelial cells results in increased secretion of several growth factors, cytokines, chemokines, and angiogenic factors, and the multifunctional angiogenic protein angiogenin is one of them. KS tissue sections were positive for angiogenin, highlighting the importance of angiogenin in KS pathogenesis. Examination of KSHV-mediated angiogenin upregulation and secretion and potential outcomes revealed that during infection of primary endothelial cells, KSHV induced a time- and dose-dependent increase in angiogenin gene expression and protein secretion beginning as early as 8 h postinfection and lasting until the fifth day of our observation period. TIVE latently transformed cells (TIVE-LTC) latently infected with KSHV secreted high levels of angiogenin. Angiogenin was also detected in BCBL-1 cells (human B cells) carrying KSHV in a latent state. Significant induction of angiogenin was observed in cells expressing KSHV ORF73 (LANA-1; latent) and ORF74 (lytic) genes alone, and moderate induction was seen with the lytic KSHV ORF50 gene. Angiogenin bound to surface actin, internalized in a microtubule-independent manner, and translocated into the nucleus and nucleolus of infected cells. In addition, it increased 45S rRNA gene transcription, antiapoptosis, and proliferation of infected cells, thus demonstrating the multifunctional nature of KSHV-induced angiogenin. These activities were dependent on angiogenin nuclear translocation, which was inhibited by neomycin. Upregulation of angiogenin led to increased activation of urokinase plasminogen activator and generation of active plasmin, which facilitated the migration of endothelial cells toward chemoattractants, including angiogenin, and chemotaxis was prevented by the inhibition of angiogenin nuclear translocation. Treatment of KSHV-infected cell supernatants with antiangiogenin antibodies significantly inhibited endothelial tube formation, and inhibition of nuclear translocation of angiogenin also blocked the expression of KSHV-induced vascular endothelial growth factor C. Collectively, these results strongly suggest that by increasing infected endothelial cell 45S rRNA synthesis, proliferation, migration, and angiogenesis, KSHV-induced angiogenin could be playing a pivotal role in the pathogenesis of KSHV infection, including a contribution to the angioproliferative nature of KS lesions. Our studies suggested that LANA-1 and vGPCR play roles in KSHV-induced angiogenesis and that the angiogenic potential of vGPCR might also be due to its ability to induce angiogenin.
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Jones, Brielle, Chaoyang Li, Min Sung Park, Anne Lerch, Vimal Jacob, Nicholas Johnson, Jin-Qiang Kuang, Sandeep Dhall, and Malathi Sathyamoorthy. "Comprehensive Comparison of Amnion Stromal Cells and Chorion Stromal Cells by RNA-Seq." International Journal of Molecular Sciences 22, no. 4 (February 14, 2021): 1901. http://dx.doi.org/10.3390/ijms22041901.

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Mesenchymal stromal cells derived from the fetal placenta, composed of an amnion membrane, chorion membrane, and umbilical cord, have emerged as promising sources for regenerative medicine. Here, we used next-generation sequencing technology to comprehensively compare amniotic stromal cells (ASCs) with chorionic stromal cells (CSCs) at the molecular and signaling levels. Principal component analysis showed a clear dichotomy of gene expression profiles between ASCs and CSCs. Unsupervised hierarchical clustering confirmed that the biological repeats of ASCs and CSCs were able to respectively group together. Supervised analysis identified differentially expressed genes, such as LMO3, HOXA11, and HOXA13, and differentially expressed isoforms, such as CXCL6 and HGF. Gene Ontology (GO) analysis showed that the GO terms of the extracellular matrix, angiogenesis, and cell adhesion were significantly enriched in CSCs. We further explored the factors associated with inflammation and angiogenesis using a multiplex assay. In comparison with ASCs, CSCs secreted higher levels of angiogenic factors, including angiogenin, VEGFA, HGF, and bFGF. The results of a tube formation assay proved that CSCs exhibited a strong angiogenic function. However, ASCs secreted two-fold more of an anti-inflammatory factor, TSG-6, than CSCs. In conclusion, our study demonstrated the differential gene expression patterns between ASCs and CSCs. CSCs have superior angiogenic potential, whereas ASCs exhibit increased anti-inflammatory properties.
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Suku, Meenakshi, Ashang Luwang Laiva, Fergal J. O’Brien, and Michael B. Keogh. "Anti-Ageing Protein β-Klotho Rejuvenates Diabetic Stem Cells for Improved Gene-Activated Scaffold Based Wound Healing." Journal of Personalized Medicine 11, no. 1 (December 22, 2020): 4. http://dx.doi.org/10.3390/jpm11010004.

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Skin wounds can lead to serious morbidity complications in diabetic patients due to the reduced healing potential of autologous stem cells. One reason for the low functional potency of stem cells from diabetic patients (diabetic stem cells) is attributed to their senescent-like nature. Here, we investigated if an anti-ageing protein, β-klotho, could be used to rejuvenate diabetic stem cells and to promote pro-angiogenic gene-activated scaffold (GAS)-induced functional response for wound healing applications. Human stem cells derived from the adipose tissue (adipose-derived stem cells (ADSCs)) of normal and diabetic (type 2) donors were used for the study. We report that the β-klotho priming facilitated inflammatory signal pruning by reducing interleukin-8 release by more than half while concurrently doubling the release of monocyte chemoattractant protein-1. Additionally, β-klotho priming enhanced the pro-angiogenic response of diabetic ADSCs on GAS by dampening the release of anti-angiogenic factors (i.e., pigment epithelium-derived factor, tissue inhibitor of metalloproteinase-1 and thrombospondin-1) while simultaneously supporting the expression of pro-angiogenic factors (i.e., Vascular Endothelial Growth Factor (VEGF), angiopoietin-2 and angiogenin). Finally, we show that β-klotho pre-treatment expedites the cellular expression of matrix proteins such as collagen IV and collagen VI, which are implicated in tissue maturation. Taken together, our study provides evidence that the synergistic effect of the pro-angiogenic GAS and β-klotho activation effectively accelerates the functional development of diabetic ADSCs for wound healing applications.
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Asosingh, Kewal, Hendrik De Raeve, Mark de Ridder, Guy A. Storme, Angelo Willems, Ivan Van Riet, Benjamin Van Camp, and Karin Vanderkerken. "Role of the Hypoxic Bone Marrow Microenvironment in Multiple Myeloma Tumor Progression." Blood 104, no. 11 (November 16, 2004): 2348. http://dx.doi.org/10.1182/blood.v104.11.2348.2348.

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Abstract Recently we reported that pre-clinical myeloma disease progression in the 5T2MM mouse model is characterized by predominant CD45+ MM-cells in the early, pre-angiogenic stage stage of slow tumor progression, followed by expansion of CD45− MM-cells during the subsequent angiogenic stage of progressive tumor growth. Unlike other cancer cells, multiple myeloma (MM) cells have to survive and to grow in a microenvironment which is already hypoxic by nature. This hypoxic bone marrow (BM) microenvironment is essential for normal hematopoiesis. However, the role of BM hypoxia in myeloma tumor progression is not known. Herein we addressed this topic in the 5T2MM mouse model. Flow cytometric analysis of control mice and 5T2MM diseased mice injected with pimonidazole hypoxyprobe indicated that both normal BM and myeloma infiltrated BM are hypoxic. However, in myelomatous BM the hypoxia was significantly decreased. Analysis of HIF-1a expression, a surrogate marker of hypoxia, by flow cytometry also demonstrated significantly lower levels of hypoxia in myeloma infiltrated BM. HIF-1a expression was found in 5T2MM-cells and was significantly higher compared to the non-tumor cell fraction. In vitro culturing of 5T2MM cells under hypoxic conditions, indicated increased activation of apoptosis inducing caspase-3 in the CD45− MM-fraction, but not in the CD45+ 5T2MM-cells, suggesting that native BM hypoxia selects the tumor population for tumor initiating CD45+ 5T2MM-cells. Although angiogeneic switch and angiogeneic heterogeneity has been reported in MM, the role of myeloma associated angiogensis is remains unclear. The decreased hypoxia in myeloma infiltrated BM adds strength to the hypothesis that myeloma associated neovascularization is functional by increasing BM oxygenation. The data also suggest that the angiogenesis allows expansion of CD45− 5T2MM-cells by decreasing BM hypoxia. All together, these findings suggest an important role of BM hypoxia in myeloma tumor progression.
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Fett, James W., Daniel J. Strydom, Roy R. Lobb, Edward M. Alderman, J. Lemuel Bethune, James F. Riordan, and Bert L. Vallee. "Isolation and characterization of angiogenin, an angiogenic protein from human carcinoma cells." Biochemistry 24, no. 20 (September 1985): 5480–86. http://dx.doi.org/10.1021/bi00341a030.

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Paul, Santanu, Steven L. Allen, Kanti R. Rai, and Nicholas Chiorazzi. "Expression of Angiogenin in Normal B Lymphocytes and B-CLL Cells." Blood 106, no. 11 (November 16, 2005): 1188. http://dx.doi.org/10.1182/blood.v106.11.1188.1188.

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Abstract Angiogenesis is critical for the clinical progression of hematopoietic malignancies and depends on a series of angiogenic factors. Angiogenin is a potent angiogenic factor produced by the host microenvironment and certain neoplastic cells. The association between angiogenin, cancer progression and poor outcome in solid tumors has been documented, but its significance in B-CLL has not been defined. A previous study suggests that B-CLL patients in Rai stage O with higher angiogenin levels have a better clinical outcome. This is surprising in light of the association of angiogenin levels with progression in solid tumors. Therefore we analyzed angiogenin mRNA and protein expression by the leukemic cells of B-CLL patients in order to correlate the production of this molecule with disease progression in B-CLL. Angiogenin was expressed in B-CLL cells as well as in B cells of normal donors, although the expression in the former was much higher than in the latter. Q PCR revealed a 7-fold induction in angiogenin-specific mRNA transcription in B-CLL cells (n=10) in comparison to normal B cells (n=13). In addition, angiogenin protein was identified by confocal microscopy both within and on the cell surface of B-CLL cells. All B-CLL cases, regardless of IgV gene mutation status, express angiogenin as defined by FACS. Approximately 85% of the cells that comprise B-CLL clones (n=28) display angiogenin on their surface membranes (range: 55-98%). Furthermore approximately 13% of polyclonal B cells from normal subjects (n=12) also produce angiogenin (range: 1–33%). Using enzyme immunoassay, we also measured serum angiogenin levels and detected significant differences in 66 B-CLL patients (median: 381 ng/mL; range: 185–875) versus 24 age- and sex-matched healthy controls (median: 301 ng/mL; range: 192–536) (P = 0.0056). Surprisingly, there was no correlation between surface and serum angiogenin levels and the different V gene-defined B-CLL subgroups. Our results show for the first time that a small fraction of normal B cells and all cases of B-CLL express angiogenin transcripts as well as intracellular and surface membrane protein. Angiogenin levels do not correlate with IgV gene mutations status or expression of surface membrane CD38. The role of angiogenin in the pathogenesis and progression of B-CLL needs further study.
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Komici, Klara, Isabella Gnemmi, Claudia Sangiorgi, Fabio Luigi Massimo Ricciardolo, Mauro Rinaldi, Antonino Di Stefano, and Ermanno Eleuteri. "Indexes of Angiogenic Activation in Myocardial Samples of Patients with Advanced Chronic Heart Failure." Medicina 55, no. 12 (November 29, 2019): 766. http://dx.doi.org/10.3390/medicina55120766.

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Background and objectives: Ischemic and idiopathic heart failure are characterized by reactive cardiac fibrosis and impaired vasculogenesis involving pro-angiogenic factors such as angiogenin, angiopoietin-1 (Ang-1), and angiopoietin-2 (Ang-2), as demonstrated in experimental models of heart failure. However, differences in the molecular pathways between these cardiomyopathies are still unclear. In this short communication, we evaluate and compare the expression of pro-angiogenic molecules in the heart tissue of patients with advanced chronic heart failure (CHF) of ischemic vs. nonischemic etiology. Materials and Methods: We obtained heart tissue at transplantation from left ventricular walls of 16 explanted native hearts affected by either ischemic (ICM) or nonischemic dilated cardiomyopathy (NIDCM). Tissue samples were examined using immunohistochemistry for angiogenic molecules. Results: We found immunopositivity (I-pos) for angiopoietin-1 mainly in the cardiomyocytes, while we observed I-pos for Ang-2 and Tie-2 receptor mainly in endothelial cells. Expression of Procollagen-I (PICP), angiogenin, Ang-1, and Tie-2 receptor was similar in ICM and NIDCM. In contrast, endothelial immunopositivity for Ang-2 was higher in ICM samples than NIDCM (p = 0.03). Conclusions: In our series of CHF heart samples, distribution of Ang-1 and angiogenin was higher in cardiomyocytes while that of Ang-2 was higher in endothelial cells; moreover, Ang-2 expression was higher in ICS than NIDCM. Despite the small series examined, these findings suggest different patterns of angiogenic stimulation in ICM and NIDCM, or at least a more altered endothelial integrity in ICD. Our data may contribute to a better understanding of the angiogenesis signaling pathways in CHF. Further studies should investigate differences in the biochemical processes leading to heart failure.
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Schanz, Andrea, Margarete Lukosz, Alexandra P. Hess, Dunja M. Baston-Büst, Jan S. Krüssel, and Christian Heiss. "hCG stimulates angiogenic signals in lymphatic endothelial and circulating angiogenic cells." Journal of Reproductive Immunology 110 (August 2015): 102–8. http://dx.doi.org/10.1016/j.jri.2015.01.011.

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Bretschneider, Henriette, Mandy Quade, Anja Lode, Michael Gelinsky, Stefan Rammelt, Stefan Zwingenberger, Klaus-Dieter Schaser, and Corina Vater. "Characterization of Naturally Occurring Bioactive Factor Mixtures for Bone Regeneration." International Journal of Molecular Sciences 21, no. 4 (February 19, 2020): 1412. http://dx.doi.org/10.3390/ijms21041412.

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In this study, the bone-regenerative potential of bioactive factors derived from adipose tissue, platelet-rich plasma (PRP) and conditioned medium from hypoxia-treated human telomerase immortalized bone-marrow-derived mesenchymal stem cells (hTERT-MSC) was investigated in vitro with the aim to develop cost-effective and efficient bone substitutes for optimized regeneration of bone defects. Adipose tissue was harvested from human donors undergoing reconstructive surgery, and adipose tissue extract (ATE) was prepared. Platelet lysates (PL) were produced by repeated freeze-thaw cycles of PRP, and hypoxia-conditioned medium (HCM) was obtained by culturing human telomerase immortalized bone-marrow-derived mesenchymal stromal cells for 5 days with 1% O2. Besides analysis by cytokine and angiogenesis arrays, ELISA was performed. Angiogenic potential was investigated in cocultures of bone-marrow-derived (BM)-MSC and human umbilical vein endothelial cells. Multiple angiogenic proteins and cytokines were detected in all growth factor mixtures. HCM and ATE contained high amounts of angiogenin and CCL2/MCP-1, whereas PL contained high amounts of IGFBP-1. Culturing cells with HCM and ATE significantly increased specific ALP activity of BM-MSC as well as tubule length and junctions of endothelial networks, indicating osteogenic and angiogenic stimulation. To achieve a synergism between chemoattractive potential and osteogenic and angiogenic differentiation capacity, a combination of different growth factors appears promising for potential clinical applications.
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Golat, Brian T., and Don F. Cameron. "Sertoli Cells Enhance Formation of Capillary-Like Structures in Vitro." Cell Transplantation 17, no. 10-11 (October 2008): 1135–44. http://dx.doi.org/10.3727/096368908787236512.

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Sertoli cells isolated from the testis (referred to as extratesticular Sertoli cells) have been shown to facilitate allo- and xenogeneic cell transplantations. It appears likely that the ability of these cells to enhance the success of cell engraftment is due, in part, to the retention of their intratesticular functions of trophic support and immunoprotection. Sertoli cells also are involved in the regulation of angiogenesis in the testis, which may also contribute to enhanced cell engraftment success facilitated by extratesticular Sertoli cells. Because the maintenance of the cell's intratesticular angiogenic function has not yet been evaluated for extratesticular Sertoli cells, this study examined the cell's ability to enhance angiogenesis in vitro. Sertoli cell conditioned media were derived from isolated rat Sertoli cell cultures and used in a rat aortic model of induced angiogenesis, in endothelial and smooth muscle cell monocultures, and in endothelial smooth muscle cocultures. An angiogenic rat cytokine array identified angiogenic factors in the control and conditioned media. Aorta sections incubated with Sertoli cell conditioned media showed a marked increase in the formation of capillary-like structures when compared to controls. Likewise, endothelial cells incubated in conditioned media organized into capillary-like structures not observed when incubated in control media. In coculture, smooth muscle cells were associated with endothelial cell-derived capillary-like structures only when incubated in conditioned media. Cytokine arrays indicated the presence and a qualitative increase of specific angiogenic growth factors in Sertoli cell conditioned media not observed in control media. Results indicate that extratesticular Sertoli cells retain their intratesticular angiogenic function in vitro.
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Dissertations / Theses on the topic "Angiogenic cells"

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A, Ahern Megan, Black Claudine P, Seedorf Gregory J, Baker Christopher D, and Shepherd Douglas P. "Hyperoxia impairs pro-angiogenic RNA production in preterm endothelial colony-forming cells." AMER INST MATHEMATICAL SCIENCES-AIMS, 2017. http://hdl.handle.net/10150/626103.

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Disruptions in the response of endothelial progenitor cells to changes in oxygen environment may present a possible mechanism behind multiple pediatric pulmonary disease models, such as bronchopulmonary dysplasia. Using high-throughput fixed single-cell protein and RNA imaging, we have created "stop-motion" movies of Thymosin. 4 (T beta 4) and Hypoxia Inducible Factor 1 alpha (HIF-1 alpha) protein expression and vascular endothelial growth factor (vegf) and endothelial nitric oxide synthase (eNOS) mRNA in human umbilical cord-derived endothelial colony-forming cells (ECFC). ECFC were grown in vitro under both room air and hyperoxia (50% O-2). We find elevated basal T beta 4 protein expression in ECFC derived from prematurely born infants versus full term infants. T beta 4 is a potent growth hormone that additionally acts as an actin sequestration protein and regulates the stability of HIF-1 alpha. This basal level increase of T beta 4 is associated with lower HIF1 alpha nuclear localization in preterm versus term ECFC upon exposure to hyperoxia. We find altered expression in the pro-angiogenic genes vegf and eNOS, two genes that HIF-1 alpha acts as a transcription factor for. This provides a potential link between a developmentally regulated protein and previously observed impaired function of preterm ECFC in response to hyperoxia.
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Foroni, Laura <1978&gt. "Resident angiogenic mesenchymal stem cells from multiorgan donor thoracic aortas." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2008. http://amsdottorato.unibo.it/976/.

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Stem cells are one of the most fascinating areas of biology today, and since the discover of an adult population, i.e., adult Stem Cells (aSCs), they have generated much interest especially for their application potential as a source for cell based regenerative medicine and tissue engineering. aSCs have been found in different tissues including bone marrow, skin, intestine, central nervous system, where they reside in a special microenviroment termed “niche” which regulate the homeostasis and repair of adult tissues. The arterial wall of the blood vessels is much more plastic than ever before believed. Several animal studies have demonstrated the presence of cells with stem cell characteristics within the adult vessels. Recently, it has been also hypothesized the presence of a “vasculogenic zone” in human adult arteries in which a complete hierarchy of resident stem cells and progenitors could be niched during lifetime. Accordingly, it can be speculated that in that location resident mesenchymal stem cells (MSCs) with the ability to differentiate in smooth muscle cells, surrounding pericytes and fibroblasts are present. The present research was aimed at identifying in situ and isolating MSCs from thoracic aortas of young and healthy heart-beating multiorgan donors. Immunohistochemistry performed on fresh and frozen human thoracic aortas demonstrated the presence of the vasculogenic zone between the media and the adventitial layers in which a well preserved plexus of CD34 positive cells was found. These cells expressed intensely HLA-I antigens both before and after cryopreservation and after 4 days of organ cultures remained viable. Following these preliminary results, we succeeded to isolate mesenchymal cells from multi-organ thoracic aortas using a mechanical and enzymatic combined procedure. Cells had phenotypic characteristics of MSC i.e., CD44+, CD90+, CD105+, CD166+, CD34low, CD45- and revealed a transcript expression of stem cell markers, e.g., OCT4, c-kit, BCRP-1, IL6 and BMI-1. As previously documented using bone marrow derived MSCs, resident vascular wall MSCs were able to differentiate in vitro into endothelial cells in the presence of low-serum supplemented with VEGF-A (50 ng/ml) for 7 days. Under the condition described above, cultured cells showed an increased expression of KDR and eNOS, down-regulation of the CD133 transcript, vWF expression as documented by flow cytometry, immunofluorescence, qPCR and TEM. Moreover, matrigel assay revealed that VEGF induced cells were able to form capillary-like structures within 6 hours of seeding. In summary, these findings indicate that thoracic aortas from heart-beating, multi-organ donors are highly suitable for obtaining MSCs with the ability to differentiate in vitro into endothelial cells. Even though their differentiating potential remains to be fully established, it is believed that their angiogenic ability could be a useful property for allogenic use. These cells can be expanded rapidly, providing numbers which are adequate for therapeutic neovascularization; furthermore they can be cryostored in appropriate cell banking facilities for later use.
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Milewski, Michael Edward. "The Angiogenic effect of Erythropoietin on Stem Cells In-Vitro." Master's thesis, Temple University Libraries, 2011. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/127595.

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Biology
M.S.
Angiogenesis is a normal and vital process that occurs during growth and development. Repair of bony defects, whether in the craniofacial complex or the alveolus, require an alloplastic or xenoplastic bone graft with angiogenic potential. This angiogenic potential is derived from existing blood vessels adjacent to the graft site. Improving the endogenous angiogenic potential with a molecule would drastically improve the survival rate of the bone graft material. This study was conducted to test the hypothesis that specific stem cell lines treated with erythropoietin, a positive promoter of angiogenesis, may increase the erythropoietin receptor expression in-vitro. In addition, this study also evaluated the vascular branching in vitro of human umbilical vein-derived endothelial cells treated with erythropoietin in the matrigel assay. Human umbilical vein-derived endothelial cells were treated for seven days with four concentrations of erythropoietin and cellular branching was evaluated in the matrigel assay. human bone marrow-derived mesenchymal stem cells and multi-potent cord blood derived unrestricted stromal stem cells were treated for seven days with erythropoietin and erythropoietin receptor expression was evaluated via reverse transcriptase real time polymerase chain reaction and real time polymerase chain reaction assays. The results of this study indicate that: erythropoietin had no effect on human umbilical vein-derived endothelial cells in the matrigel assay from a qualitative perspective, after treating multi-potent cord blood derived unrestricted stromal stem cells cells for 7 days with erythropoietin, there was no statistically significant difference between treatment groups when compared to control, and after treating human bone marrow-derived mesenchymal stem cells cells for 7 days with erythropoietin, the 20 U/ml treatment group showed a statistically significant reduction of the erythropoietin receptor as compared to the control group.
Temple University--Theses
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Latham, Nicholas. "Second Generation Cardiac Cell Therapy: Combining Cardiac Stem Cells and Circulating Angiogenic Cells for the Treatment of Ischemic Heart Disease." Thèse, Université d'Ottawa / University of Ottawa, 2013. http://hdl.handle.net/10393/24293.

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Blood-derived circulatory angiogenic cells (CACs) and resident cardiac stem cells (CSCs) have both been shown to improve cardiac function after myocardial infarction (MI) but the superiority of either cell type has long been an area of speculation with no definitive head-to-head trial. In this study, we compared the paracrine profile of human CACs and CSCs, alone or in combination. We characterized the therapeutic ability of these cells to salvage myocardial function in an immunodeficient mouse model of MI by transplanting these cells as both single and dual cell therapies seven days after experimental anterior wall MI. CACs and CSCs demonstrated unique paracrine repertoires with equivalent effects on angiogenesis, stem cell migration and myocardial repair. Combination therapy with both cell types synergistically improves post infarct myocardial function greater than either therapy alone. This synergy is likely mediated by the complementary paracrine signatures that promote revascularization and the growth of new myocardium.
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Giordano, Céline. "Pre-Clinical Evaluation of Biopolymer Delivered Circulating Angiogenic Cells in Hibernating Myocardium." Thèse, Université d'Ottawa / University of Ottawa, 2011. http://hdl.handle.net/10393/20619.

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Vasculogenic cell-based therapy combined with tissue engineering is a promising revascularization strategy for patients with hibernating myocardium, a common clinical condition. We used a clinically relevant swine model of hibernating myocardium to examine the benefits of biopolymer-supported delivery of circulating angiogenic cells (CACs) in this context. Twenty-five swine underwent placement of an ameroid constrictor on the left circumflex artery (LCx). After 2 weeks, positron emission tomography measures of myocardial blood flow (MBF) and myocardial flow reserve (MFR) were reduced in the affected region (both p<0.001). Hibernation (mismatch) was specific to the LCx territory. Swine were randomized to receive intramyocardial injections of PBS control (n=10), CACs (n=8), or CACs + a collagen-based matrix (n=7). At follow-up, stress MBF and MFR were increased only in the cells+matrix group (p<0.01), and mismatch was lower in the cells+matrix treated animals (p=0.02) compared to controls. Similar results were found using microsphere-measured MBF. Wall motion abnormalities and ejection fraction improved only in the cells+matrix group. This preclinical swine model demonstrated ischemia and hibernation, which was improved by the combined delivery of CACs and a collagen-based matrix. To our knowledge, this is the first demonstration of the mechanisms and effects of combining progenitor cells and biopolymers in the setting of myocardial hibernation, a common clinical condition in patients with advanced coronary artery disease.
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Ward, K. L. "Analysing the angiogenic potential of mesothelial cells in vitro and in vivo." Thesis, University of Liverpool, 2017. http://livrepository.liverpool.ac.uk/3008101/.

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Webster, Katie Elizabeth. "Angiogenesis in endometriosis : the role of circulating angiogenic cells and the endometrium." Thesis, University of Oxford, 2012. http://ora.ox.ac.uk/objects/uuid:33c8921c-8320-4559-8298-52d85c8a2987.

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Endometriosis is a common cause of subfertility and pelvic pain, affecting up to 10% of women of reproductive age. It is characterised by the presence of endometrial-like tissue outside the uterus. The development of the disease is still poorly understood and, currently, the diagnosis relies on visualisation of typical lesions during surgery. There is great interest in identifying biomarkers to assist in diagnosis and disease management. Blood vessel development is known to be a crucial feature of endometriosis, but the mechanisms involved in angiogenesis are not well described for this disease. Most vessel development relies on the proliferation and migration of pre-existing endothelial cells. However, there may also be roles for cells derived from peripheral blood (circulating angiogenic cells) and surrounding stromal cells. In this thesis, the contribution of these different cell types to vessel development in endometriosis is assessed. In chapter 2, a robust protocol was optimised to identify circulating angiogenic cells (CACs) with flow cytometry. The reliability of the protocol was verified, and the level of these cells was found not to fluctuate with the menstrual cycle in healthy women (P=0.279, F=1.359, 3 d.f.). In chapter 3, levels of CACs in women with and without endometriosis were found to be equivalent (0.0835% ± 0.0422 compared to 0.0724% ± 0.0414), demonstrating that they have no use as a disease biomarker. In chapter 4, isolation and culture of endothelial cells from the endometrium was attempted. However, a pure culture of endometrial endothelial cells could not be obtained, which may be due to contamination by other cell types or cellular transdifferentiation. Finally, in chapter 5, the contribution of endometrial stromal cells to vessel development was considered. Stromal cells were found not to differentiate towards an endothelial cell phenotype, but were able to participate in tube formation assays. However, the presence of endometriosis did not influence this behaviour.
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Zhuge, Xin. "CXCL16 is a novel angiogenic factor for human umbilical vein endothelial cells." Kyoto University, 2005. http://hdl.handle.net/2433/144481.

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Russell, Hugh. "Early angiogenic change in dental pulp stromal cells cultured on biomimetic matrices." Thesis, University of Leeds, 2016. http://etheses.whiterose.ac.uk/13395/.

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Revascularisation of the devitalised root canal is the Holy Grail of Endodontics and is being hotly pursued by many teams of clinicians but has yet to be achieved. The overall aim of this work was to attempt to induce early angiogenesis in human dental pulp stromal cells (DPSCS) in vitro and in vivo using a biomimetic approach based on combining scaffolds comprised of ECM components with DPSCs as a first step towards a tissue engineering strategy for dental pulp regeneration. After isolating DPSCs using collagenase digest, they were cultured on 1% hyaluronic acid (HyA) or Types I and III collagen matrices used either singly or in combination to determine the ability of these scaffolds to support/induce early angiogenic change in DPSCs both in vitro and in vivo. Angiogenic change was determined using a combined approach of DNA quantification, histology, immunohistochemistry to detect the angiogenic markers CD31 and CD34 and quantitative RT-PCR. DPSCs were shown to attach and proliferate on Type I and III collagen membranes in vitro but early angiogenic change in vitro was evidenced only when 1% HyA gel was used, including in the absence of the morphogen rhVEGF165 as shown immunohistochemically. PCR at two and five days post-seeding showed an up-regulation of CD31 and CD34 genes dependant on culture conditions, with CD31 being upregulated early and CD34 later in the culture period. A modified tooth slice model containing a combination of HyA/collagen scaffold/DPSC constructs within its lumen also showed positive early angiogenic change in vitro. SEM examination further confirmed that DPSCs could attach, colonise and proliferate to/on the combined scaffold. The same combined scaffold-tooth slice model ± DPSCs used in vivo in nude mice showed cellular ingress into the lumen with a soft tissue closely resembling dental pulp-like tissues in its appearance with new tubule-like material grown on from the dentinal tubules of the tooth slice. There was a defined demarcation line between this latter material and the dentinal tubules of the tooth slice and the new material closely resembled predentine or dentine-like matrix in appearance and stained strongly for CD31 and CD34 markers. It also had a layer of cells adjacent to and in intimate contact with its deposition front, whose cell processes transited the new tubule-like material and continued into the dentine tubules of the tooth slice for some distance. Interestingly, this neo-tissue was independent of the addition of DPSCs to the construct. The results suggest that biomimetic scaffolds based upon components of the pulp extracellular matrix may provide a useful platform for future engineering of a vascularised replacement dental pulp.
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Ostojic, Aleksandra. "Use of a Collagen I Matrix to Enhance the Potential of Circulating Angiogenic Cells (CACs) for Therapy." Thesis, Université d'Ottawa / University of Ottawa, 2015. http://hdl.handle.net/10393/31931.

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Acute myocardial infarction (MI) is the end result of many cardiovascular diseases and is one of the leading causes of death in the western world. Cell therapy, using circulating angiogenic cells (CACs) or CD34+ cells from peripheral blood, is one approach under investigation for restoring blood flow and function to the ischemic heart. However, the numbers of CACs and CD34+ circulating cells are inversely proportional to the severity of cardiovascular disease and age; therefore, there is a need to increase their numbers and/or function for therapy. One possibility is to enhance the therapeutic potential of the cells with the use of a biomaterial. In this study, we used a collagen matrix to culture human CD34+ circulating cells, and evaluated the effect of the matrix on CD34+ cell properties and function. The matrix was able to successfully increase proliferation, migration, CD34+ phenotype and branching in an angiogenesis assay. These functional benefits may be associated with the sonic hedgehog (Shh) pathway. The collagen matrix was previously shown to enhance the function of healthy CACs, but its ability to do the same for CACs from coronary artery disease patients is unknown. In this study, the matrix was shown to enhance the viability, proliferation and angiogenic potential of patient CACs. Furthermore, gene expression for integrins and Shh pathway components in the sub-population of CD34+ cells was similar between patient and healthy donors when isolated from CACs. This work provides insight into the mechanisms for the observed matrix-enhanced function of therapeutic CACs and CD34+ cells from both healthy and CAD patient donors.
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Books on the topic "Angiogenic cells"

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International Workshop "Novel Angiogenic Mechanisms" (2002 Columbus, Ohio). Novel angiogenic mechanisms: Role of circulating progenitor endothelial cells. New York: Kluwer Academic/Plenum, 2003.

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I, Moldovan Nicanor, ed. Novel angiogenic mechanisms: Role of circulating progenitor endothelial cells. New York: Kluwer Academic/Plenum, 2003.

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Tran, Jennifer. Sensitizing tumor vascular endothelial cells to anti-angiogenic therapy. 2004.

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Moldovan, Nicanor I. Novel Angiogenic Mechanisms: Role Of Circulating Progenitor Endothelial Cells. Springer, 2012.

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Wang, Angela. Effect of estrogen on angiogenic factor expression in cultured uterine cells. 2005.

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Wang, Angela. Effect of estrogen on angiogenic factor expression in cultured uterine cells. 2005.

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Moldovan, Nicanor I. Novel Angiogenic Mechanisms: Role of Circulating Progenitor Endothelial Cells (Advances in Experimental Medicine and Biology). Springer, 2003.

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Weller, Michael, Michael Brada, Tai-Tong Wong, and Michael A. Vogelbaum. Astrocytic tumours: diffuse astrocytoma, anaplastic astrocytoma, glioblastoma, and gliomatosis cerebri. Oxford University Press, 2017. http://dx.doi.org/10.1093/med/9780199651870.003.0003.

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Astrocytic gliomas are primary brain tumours thought to originate from neural stem or progenitor cells. They are assigned grades II, III, or IV by the World Health Organization according to degree of malignancy as defined by histology. The following molecular markers are increasingly used for diagnostic subclassification or clinical decision-making: 1p/19q co-deletion status, O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation status, and isocitrate dehydrogenase 1 and 2 mutation status. Extent of resection is a favourable prognostic factor, but surgery is never curative. Radiotherapy prolongs progression-free survival across all astrocytic glioma entities. Alkylating agent chemotherapy is an active treatment in particular for patients with MGMT promoter-methylated tumours. Anti-angiogenic therapies have failed to improve survival, and the current focus of major clinical trials is on novel targeted agents or on immunotherapy.
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Yu, Joanne Linda. Heterogeneous vascular dependence of tumor cell subpopulations and implications for anti-angiogenic therapy. 2002.

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Connor, Thomas, and Patrick H. Maxwell. Von Hippel–Lindau disease. Edited by Neil Turner. Oxford University Press, 2015. http://dx.doi.org/10.1093/med/9780199592548.003.0332.

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Von Hippel–Lindau (VHL) disease is a dominantly inherited familial cancer syndrome caused by germline mutations in the VHL tumour suppressor gene. The most frequent manifestations of VHL disease are retinal and central nervous system haemangioblastomas, clear cell renal cell carcinomas, and phaeochromocytomas. Genetic testing and active screening for clinical manifestations is now started in childhood and has greatly improved the prognosis for patients with VHL disease. The VHL protein plays a critical role in regulating the cellular response to changes in oxygen tension. Loss of VHL function results in constitutive activation of a range of angiogenic and metabolic pathways. New drug therapies have been developed that reverse some of the cellular consequences of VHL loss of function in kidney cancer.
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Book chapters on the topic "Angiogenic cells"

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Vaughan, Erin E., and Timothy O’Brien. "Isolation of Circulating Angiogenic Cells." In Methods in Molecular Biology, 351–56. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-61779-980-8_25.

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Roitbak, Tamara. "Pro-angiogenic Properties of the Neural Stem/Progenitor Cells." In Stem Cells and Cancer Stem Cells, Volume 5, 241–48. Dordrecht: Springer Netherlands, 2012. http://dx.doi.org/10.1007/978-94-007-2900-1_23.

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Kumar, V. B. S., R. I. Viji, M. S. Kiran, and Perumana R. Sudhakaran. "Angiogenic Response of Endothelial Cells to Fibronectin." In Advances in Experimental Medicine and Biology, 131–51. New York, NY: Springer New York, 2012. http://dx.doi.org/10.1007/978-1-4614-3381-1_10.

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Noonan, Douglas M., Agostina Ventura, Antonino Bruno, Arianna Pagani, and Adriana Albini. "The Angiogenic Switch: Role of Immune Cells." In Immunologic Signatures of Rejection, 57–75. New York, NY: Springer New York, 2010. http://dx.doi.org/10.1007/978-1-4419-7219-4_5.

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Ribatti, Domenico. "Mast Cells in Angiogenesis: The Role of Angiogenic Cytokines." In Biochemical Basis and Therapeutic Implications of Angiogenesis, 157–67. Cham: Springer International Publishing, 2017. http://dx.doi.org/10.1007/978-3-319-61115-0_8.

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Seandel, Marco, Andrea T. Hooper, and Shahin Rafii. "Contribution of Endothelial Progenitor Cells to the Angiogenic Process." In Angiogenesis, 239–48. Boston, MA: Springer US, 2008. http://dx.doi.org/10.1007/978-0-387-71518-6_21.

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Ribatti, Domenico, and Angelo Vacca. "Role of Endothelial Cells and Fibroblasts in Multiple Myeloma Angiogenic Switch." In Plasma Cell Dyscrasias, 51–61. Cham: Springer International Publishing, 2016. http://dx.doi.org/10.1007/978-3-319-40320-5_5.

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Ostojic, Aleksandra, Suzanne Crowe, Brian McNeill, Marc Ruel, and Erik J. Suuronen. "Preparation and Characterization of Circulating Angiogenic Cells for Tissue Engineering Applications." In Methods in Molecular Biology, 27–38. New York, NY: Springer New York, 2014. http://dx.doi.org/10.1007/978-1-4939-1047-2_3.

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Loffredo, Stefania, Rosaria Ilaria Staiano, Francescopaolo Granata, Arturo Genovese, and Gianni Marone. "Immune Cells as a Source and Target of Angiogenic and Lymphangiogenic Factors." In Chemical Immunology and Allergy, 15–36. Basel: S. KARGER AG, 2013. http://dx.doi.org/10.1159/000353316.

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Vu, Ngoc Bich, Ha Thi-Ngan Le, Thuy Thi-Thanh Dao, Lan Thi Phi, Ngoc Kim Phan, and Van Thanh Ta. "Allogeneic Adipose-Derived Mesenchymal Stem Cell Transplantation Enhances the Expression of Angiogenic Factors in a Mouse Acute Hindlimb Ischemic Model." In Stem Cells: Biology and Engineering, 1–17. Cham: Springer International Publishing, 2017. http://dx.doi.org/10.1007/5584_2017_63.

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Conference papers on the topic "Angiogenic cells"

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Sheikh, Abdul Q., Toloo Taghian, Andrei Kogan, and Daria A. Narmoneva. "Electric Field Stimulates Angiogenesis via Activation of MAPK/ERK Signaling in Microvascular Endothelial Cells." In ASME 2012 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2012. http://dx.doi.org/10.1115/sbc2012-80851.

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Different types of physiological amplitude (40–250 mV/mm) [1] electric fields (EFs) have been shown to influence a wide variety of biological systems [2] and have been used as a therapeutic tool for tissue repair [3]. There has been emerging evidence that certain types of EFs can promote angiogenesis and tissue vascularization [4]. Studies have shown that direct current EF and pulsed EF induced angiogenic responses including cell migration, VEGF release and cytoskeletal reorganization in human endothelial cells (HUVECs) [4, 5]. Similarly different types of EF induced the activation of intracellular MAPK pathways in several non-endothelial cell types. [6, 7]. However, the effect of EF with different modalities on endothelial angiogenic responses and the intracellular pathways remain unknown.
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Lee, Sue Hyun, Angela L. Zachman, Desirae L. Deskins, Pampee P. Young, and Hak-Joon Sung. "ROS-Responsive Scaffold for Angiogenic Differentiation of Mesenchymal Stem Cells." In ASME 2013 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/sbc2013-14553.

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Vascularization of a tissue-engineered construct enables efficient transport of nutrients and waste products; it is necessary for successful long-term tissue growth and host integration. Although significant progress has been made, sufficient vascularization of engineered constructs is still a major challenge, limiting clinical applications of tissue engineering (TE) approaches [1]. Successful vascularization promotes the interactions of TE implants with host tissues, leading to efficient tissue regeneration. Therefore, there is an unmet need to develop a more efficient method to vascularize TE constructs. In particular, obtaining a reliable source of endothelial cells (ECs) that line all blood vessels is a critical and challenging step towards successful vascularization of TE constructs, empowering TE to be applied in a larger scale and scope.
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Ryan, David T., Jingzhe Hu, Byron L. Long, and Amina A. Qutub. "Predicting Endothelial Cell Phenotypes in Angiogenesis." In ASME 2013 2nd Global Congress on NanoEngineering for Medicine and Biology. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/nemb2013-93124.

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Controlling endogenous angiogenesis, or the formation of new capillaries, is a potential therapeutic strategy for numerous diseases including cancer, stroke and cardiovascular disease (1–4). These efforts have been met with mixed success in the clinic, partly due to an inadequate understanding, and thus control, of the mechanisms that influence the endothelial cells that form capillaries (1, 3). In order to control angiogenesis in an effort to improve treatment responses, quantitative information about endothelial cell behavior must be used to build accurate models of vascular network formation. In this paper, we introduce a method to identify and classify endothelial cell responses to angiogenic stimuli through sophisticated image analysis. The presented automated image processing tools and classification framework allow for rapid quantitative investigations of cellular images. Results of our analysis demonstrate that endothelial cells can be grouped into distinct morphological phenotypes as a function of their responses to combinations of angiogenic growth factor stimuli. Information on phenotypic behavior and responses will be applied towards predicting and guiding cell behavior for therapeutic design.
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Buchanan, Cara F., Elizabeth Voigt, Christopher S. Szot, Joseph W. Freeman, Pavlos P. Vlachos, and Marissa Nichole Rylander. "Shear Stress Mediates Angiogenic Gene Expression in a Microfluidic Tumor Vascular Model." In ASME 2012 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2012. http://dx.doi.org/10.1115/sbc2012-80286.

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While research has shown that the fluid mechanics of the tumor vasculature reduce transport and uptake of therapeutics, the underlying role of these stresses in regulating tumor-endothelial cell signaling and neovascularization are not well understood. Understanding the reciprocal interaction between endothelial and tumor cells to mediate angiogenesis, and the effect of fluid shear on this process, may offer insight into the development of improved treatment modalities to control highly vascularized tumors. We have previously shown that breast cancer cells cultured under 2D, static conditions with endothelial cells significantly increase expression of pro-angiogenic factors vascular endothelial growth factor (VEGF) and angiopoietin 2 (ANG2) [1]. These preliminary results motivated the investigation of tumor-endothelial cross-talk under 3D, dynamic co-culture conditions.
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Gnanamony, Manu, Indra Mohanam, and Sanjeeva Mohanam. "Abstract 5213: RUNX2 modulates the angiogenic potential of human neuroblastoma cells." In Proceedings: AACR 106th Annual Meeting 2015; April 18-22, 2015; Philadelphia, PA. American Association for Cancer Research, 2015. http://dx.doi.org/10.1158/1538-7445.am2015-5213.

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Taraseviciene-Stewart, L., L. Slepikas, and E. Gerasimovskaya. "The Protective Role of T Cells Against Angiogenic Potential of Endothelial Cells in Pulmonary Circulation." In American Thoracic Society 2009 International Conference, May 15-20, 2009 • San Diego, California. American Thoracic Society, 2009. http://dx.doi.org/10.1164/ajrccm-conference.2009.179.1_meetingabstracts.a2352.

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Wood, Levi B., Roger D. Kamm, and H. Harry Asada. "Time Lapse Observation Based Modeling and Identification of Cell Behaviors in Angiogenic Sprout Development." In ASME 2010 Dynamic Systems and Control Conference. ASMEDC, 2010. http://dx.doi.org/10.1115/dscc2010-4139.

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This paper presents a method for deriving dynamic equations for Endothelial Cell (EC) motion and estimating parameters based on time lapse imagery of angiogenic sprout development. Angiogenesis is the process whereby a collection of endothelial cells sprout out from an existing blood vessel, degrade the surrounding scaffold and form a new blood vessel. Sprout formation requires that a collection of ECs all work together and coordinate their movements and behaviors. The process is initiated and guided by a collection of external growth factors. In addition, the individual cells communicate and respond to each other’s movements to behave in a coordinated fashion. The mechanics of cell coordination are extremely complex and include both chemical and mechanical communication between cells and between cells and the matrix. Despite the complexity of the physical system, with many variables that cannot be measured in real time, the ECs behave in a predictable manner based on just a few quantities that can be measured in real time. This work presents a methodology for constructing a set of simple stochastic equations for cell motion dependent only on quantities obtained from time lapse data observed from in vitro experiments. Model parameters are identified from time lapse data using a Maximum Likelihood Estimator.
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Alunno, A., M. Manetti, O. Bistoni, S. Cipriani, L. Ibba Manneschi, and R. Gerli. "FRI0265 Angiogenic t cells in primary sjÖgren’s syndrome: a double-edged sword." In Annual European Congress of Rheumatology, EULAR 2018, Amsterdam, 13–16 June 2018. BMJ Publishing Group Ltd and European League Against Rheumatism, 2018. http://dx.doi.org/10.1136/annrheumdis-2018-eular.3062.

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Balaji, Swathi, Sachin S. Vaikunth, Jignesh K. Parvadia, Timothy M. Crombleholme, and Daria A. Narmoneva. "In Situ Tissue Engineering Using Angiogenic Nanoscaffold Enhances Diabetic Wound Healing in db/db Mouse Model." In ASME 2008 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2008. http://dx.doi.org/10.1115/sbc2008-192198.

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Tissue engineering offers an attractive alternative for treatment of chronic nonhealing diabetic ulcers, which account for more than 27% of the $10.9 billion total diabetic health care costs in the US annually [1]. The harsh environment of a diabetic ulcer is characterized by reduced expression of angiogenic factors, insufficient vascularization, excess protease activity, matrix degradation and hyperglycemia-induced cell apoptosis [2]. A major factor contributing to insufficient neovascularization in diabetic nonhealing wounds may be deficiency in the recruitment of endothelial cells (ECs) and endothelial precursor cells (EPCs) to the wound site [3]. Recent studies focusing on altering the wound’s cellular and molecular environment using bone-marrow-derived stem cells, growth factors (delivered either directly or using gene or cell therapy), bioengineered skin constructs, and biological matrices, such as collagen and hyaluronic acid gels had promising wound healing outcomes [4]. These studies suggest that strategies aimed at modifying the extracellular environment of the diabetic wound to enhance cell survival and angiogenesis are promising for development of new therapies for diabetic wound healing.
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Kang, Hojin, Kayla J. Bayless, and Roland Kaunas. "Effects of Fluid Shear Stress on Endothelial Cell Invasion Into Three-Dimensional Matrices." In ASME 2007 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2007. http://dx.doi.org/10.1115/sbc2007-176207.

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We have previously developed a cell culture model to study the effects of angiogenic factors, such as sphingosine-1-phosphate (S1P), on the invasion of endothelial cells into the underlying extracellular matrix. In addition to biochemical stimuli, vascular endothelial cells are subjected to fluid shear stress due to blood flow. The present study is aimed at determining the effects of fluid shear stress on endothelial cell invasion into collagen gels. A device was constructed to apply well-defined fluid shear stresses to confluent human umbilical vein endothelial cells (HUVECs) seeded on collagen gels. Fluid shear stress induced significant increases in cell invasion with a maximal induction at ∼5 dyn/cm2. These results provide evidence that fluid shear stress is a significant stimulus for endothelial cell invasion and may play a role in regulating angiogenesis.
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Reports on the topic "Angiogenic cells"

1

Chung, Leland W. Accelerated Tumor Cell Death by Angiogenic Modifiers. Fort Belvoir, VA: Defense Technical Information Center, August 2003. http://dx.doi.org/10.21236/ada418654.

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Chung, Leland W., Chia-Ling Hsieh, Michael Bradley, and Mitchell H. Sokoloff. Accelerated Tumor Cell Death by Angiogenic Modifiers. Fort Belvoir, VA: Defense Technical Information Center, August 2001. http://dx.doi.org/10.21236/ada403672.

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