Utilizing Endothelial Cell and Astrocyte Co-Cultures and Improving Differentiation Efficiency for the Treatment of Peripheral Arterial Disease
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Issue Date
2014
Authors
Al-Mubarak, Husain Adel '14
Degree
MS in Biotechnology
Advisor
Mousa, Shaker
Committee Members
Davis, Paul
Al-Omran, Mohammed
Al-Omran, Mohammed
Journal Title
Journal ISSN
Volume Title
Abstract
Atherosclerosis is a progressive disease that leads to limb loss and heart failure in peripheral arterial disease and coronary artery disease respectively. A cell-based approach to improving angiogenesis focuses on optimizing the cell type and delivery method for these conditions. Our aim was to examine the effects of co-culture on improving sprout growth and epigenetic reprograming on improving endothelial differentiation yield of adipose derived Mesenchymal Stromal Cells (adMSCs). Endothelial sprout characteristics of Human Umbilical Vein Endothelial Cells (HUVECs) in 2D and 3D assays with Normal Human Astrocytes (NHAs) and adMSCs were quantified over time. A modified 2-day epigenetic reprograming protocol utilizing Valproic acid, 5'-Azacytidine, and caffeine was used prior to a standard 7-day VEGF-A endothelial differentiation regimen on adMSCs. Co-culture of HUVECs with NHAs or adMSCs resulted in longer mean tube length, prolonged growth phase, and delay of senescence in 3D sprout formation assays over 9 days of culture compared to baseline. NHAs were able to associate with HUVEC tubes at branch points and segments in 2D assays, whereas adMSCs nearly inhibited native abilities. The modified epigenetic reprograming protocol did not result in improved phenotypic or functional differentiation of adMSCs as determined with CD31 expression and tube formation ability, respectively. In conclusion, we have demonstrated the ability of normal human astrocytes and adipose derived mesenchymal stromal cells to promote the growth and survival of three dimensional endothelial sprouts over 9 days. Additionally, we demonstrated the ability of normal human astrocytes to associate with endothelial cells in 2D tubes in an enabling manner. Our attempt at improving endothelial cell differentiation of adipose derived mesenchymal stromal cells by our modified epigenetic reprograming protocol was unsuccessful. However, it did produce clusters of cells that had morphological resemblance to pluripotent cells.
Citation
Al-Mubarak, Husain. "Utilizing Endothelial Cell and Astrocyte Co-Cultures and Improving Differentiation Efficiency for the Treatment of Peripheral Arterial Disease." Albany College of Pharmacy and Health Sciences, New York, Proquest/UMI, 2014.
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