Self-renewal and Proliferation of Murine Embryonic Stem Cells:A Study of Glycosaminoglycans Effect on Feeder-Free Cultures

摘要

The self-renewal and proliferation of murine embryonic stem(ES)cells can be preserved indefinitely in the presence of the mouse embryonic fibroblast(MEF)feeder layer.Since the feeder layer has several drawbacks,including viral contamination and large-scale production,gelatin solutions are the most prominent alternative to replacing it.In this investigation,self-renewal and proliferation of ES cells was carried out by supplementing the gelatin solution with glycosaminoglycan components,such as chondroitin sulfate and heparin.The methylcellulose-based embryoid body(EB)assay was used to evaluate the self-renewal ability of the ES cells.Chondroitin sulfate and heparin were mixed with 0.1% gelatin solution in 0:100,50:50 and 100:0(heparin/chondroitin sulfate)ratios and compared with 0.1% gelatin(positive control)and MEF(negative control)in three independent parallel experiments.Chondroitin sulfate addition to 0.1% gelatin enhanced the cell population and the number of EBs by 57 and 32%,respectively.