中山大学学报（医学科学版）2012，Vol.33Issue(6)：828-834,847,8.
无动物源性成分人胚胎干细胞培养体系与其它人胚胎干细胞培养体系的比较
Comparison of A Novel Xeno-free Culture System with Other Different Culture Systems for HESC
摘要
Abstract
[Objective] By comparing the undifferentiated and pluripotent marks of hESC in four different culture systems to evaluate and promote more optimal hESC culture environment. [Methods] The four culture systems consist of xeno-free human foreskin fibroblasts (XF-HFF) combined with chemically-defined medium (CDM) (XF-HFF/CDM group), XF-HFF combined with 200 mL/L HS medium (XF-HFF/HS group), HS-matrix coated on plates without feeder layers combined with CDM (HS-matrix/CDM group), and xeno-contained human foreskin fibroblasts (XC-HFF) combined with 200 mL/L KSR medium (XC-HFF/KSR group).The culture efficiencies were evaluated by morphology, cell growth curve, flow cytometry and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) among these culture systems. Further more, we characterized the sternness of hESC after long-term culture in XF-HFF/CDM culture system. [Results] The rates of characteristic surface undifferentiated markers (SSEA4, Tral-60) and proliferate factors (hTERT, Oct3/4) expression of hESC cultured in XF-HFF/CDM were significantly higher than which in XF-HFF/ HS and in HS-matrix/CDM(P < 0.01) , but were comparable to which in XC-HFF/KSR. After long-term cultured in XF-HFF/CDM, hESC still expressed surface undifferentiated markers of SSEA3, SSEA4 and pluripotent markers of hTERT, Oct3/4, and maintained pluripotency and karyotype stability, over 40 passages. [ Conclusion ] The new xeno-free culture conditions demonstrated similar culture efficiency to KSR medium with HFF, and were more effective than CDM with HS-matrix, or HS medium with HFF.关键词
人胚胎干细胞/无外源性成分/成分确定/比较/培养Key words
human embryonic stem cell/ xeno-free/ chemically-defined medium/ comparison/ culture分类
医药卫生引用本文复制引用
张丹,麦庆云,李涛,徐艳文,丁晨辉,宏苹苹,曾艳红,周灿权..无动物源性成分人胚胎干细胞培养体系与其它人胚胎干细胞培养体系的比较[J].中山大学学报（医学科学版）,2012,33(6):828-834,847,8.基金项目
国家"863"项目(2005AA219010) （2005AA219010）
国家"973"项目(2007CB948103) （2007CB948103）
