BIOT 324 |
| We report the development and characterization of a novel microporous membrane-based indirect co-culture system, which physically separates human embryonic stem cells (hESCs) from the feeder layer, while allowing real time conditioning of the medium, and serves as an attractive alternative to direct co-culture feeder layer-dependent propagation. Experimental outcomes demonstrate the potential of this propagation system in maintaining the undifferentiated state of two hESC lines (BG01v and WA09) in prolonged culture. The hESCs cultured is this system not only demonstrated expression of several pluripotent characteristics, but also exhibited global gene expression profiles similar to hESCs propagated using other routine hESC passaging techniques. Additionally, we demonstrate that a completely 'human' expansion method is possible based on prolonged culture of hESCs in indirect co-culture with human fibroblasts. These results represent a significant development in properly segregating hESCs from their feeders, thus eliminating cell mixing, contamination, and providing the cells with a superior environment for real-time nourishment and controlled self-renewal. Overall, this development in hESC propagation could have wide-reaching applications for self-renewal and differentiation studies within the field of stem cell biology. |
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Stem Cells: Engineering the Embryonic and Adult Stem Cell Niche
8:30 AM-11:05 AM, Wednesday, August 20, 2008 Philadelphia Marriott -- Franklin 8, Oral
Division of Biochemical Technology |