Human embryonic stem cells (hESCs) can provide a unique approach for novel tissue engineering applications. Previous groups have shown that hESCs can differentiate into specialized cell types through the generation of human embryoid bodies (hEBs). These multi-cellular constructs are then subjected to suspension culture for several weeks. Traditional hESC differentiation techniques have yielded non-homogeneous EBs derived in standard static cultures providing an inefficient platform for cellular viability and embryonic modeling. Here, our study aimed at systematically comparing the formation, growth, and differentiation capabilities of hESC-derived hEBs in dynamic and static suspension cultures. We used a continuous flow perfusion slow turning lateral vessel, STLV, system (Synthecon) to model after an in vivo environment. This study is in part of a larger study investigating the role of HOXB5 in the human endothelial differentiation pathway. Embryoid bodies were created by hanging drops and then subjected to static or dynamic culture for 10 days. Cells were harvested and a simple Alkaline Phosphatase assay was used to determine if the system was viable for propagating hEB. We show that the STLV system is viable for our future studies and this system more efficient at maintaining hEBs.
| Identifer | oai:union.ndltd.org:vcu.edu/oai:scholarscompass.vcu.edu:etd-2665 |
| Date | 12 January 2009 |
| Creators | Collier, Claiborne |
| Publisher | VCU Scholars Compass |
| Source Sets | Virginia Commonwealth University |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Theses and Dissertations |
| Rights | © The Author |
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