无动物源性培养系统对人胚胎干细胞生长的支持作用

目的:建立完全非动物源性培养系统,为人胚胎干细胞定向诱导为各系统细胞应用于临床、消除动物源性污染打下基础。方法:分别采用人包皮成纤维细胞、胎儿肌肉细胞及人子宫内膜基质细胞为饲养层细胞,以血清替代品取代动物血清,支持人胚胎干细胞生长,观察其增殖和分化情况。结果:人包皮成纤维细胞、胎儿肌肉细胞均能很好支持人胚胎干细胞生长增殖,并保持90%左右未分化表型:AKP染色强阳性,SSEA-4、TRA-1-81表达强阳性,可见OCT-4 mRNA表达,核型正常。人子宫内膜基质细胞不能支持hESC的生长。结论:完全可以使用非动物源性培养系统支持人胚胎干细胞增殖,从而为胚胎干细胞定向诱导分化应用于临床打下坚实基础

Supporting effect of non -animal contamination culture system on human embryonic stem cells growth

Objective:To establish a non-animal contamination culture system for clinical apply of human embryonic stem cells(hESC).Methods:Human foreskin fibroblast cell,fetal muscle cell and human endometrium stromal cell were used as feeder layer,FBS was replaced with serum replacement,then hESC in such a system were cultured,proliferation and differentiation characteristics of hESC were detected.Results:Foreskin fibroblast cell,fetal muscle cell could successfully support the hESC proliferation and 90% hESC maintained undifferented phenotype: positive staining of AKP,SSEA-4 and TRA-1-81,expression of OCT-4 mRNA and hESC with normal karotype.Human endometrium couldn't support the growth of hESC.Conclusion:Non-animal contamination culture system can completely support hESC growth,which provides a solid foundation for clinical apply of hESC and its derivations.