不同培养体系下iPS细胞分化为造血干细胞差异的研究

Effects of Different Culture Systems on the Hematopoietic Differentiation Ability of iPS Cells

目的:探讨不同培养体系条件下诱导多能干细胞(iPS)体外分化为造血干细胞的能力差异。方法:比较2种无滋养层培养液——E8和mTESR及经典ES培养体系下培养的iPS细胞,在体外与小鼠骨髓基质细胞OP9细胞共培养,诱导iPS细胞分化为造血干/祖细胞。采用流式细胞术及实时定量PCR方法分别检测造血特异标志物的表达,比较3种培养液对iPS体外造血分化的影响。结果:3种不同培养液培养的iPS均可分化为造血干细胞,经典ES培养体系培养的iPS诱导造血效率达28.4%,明显高于无滋养层E8和m TESR培养体系。结论:iPS与OP9共培养可分化为造血干/祖细胞,且使用经典ES培养液培养iPS更有利于其向造血系的分化。

Objective：To investigate the in vitro effects of different culture systems on hematopoietic differentiation ability of induced pluripotent stem（iPS） cells.Method：Two culture systems including E8 and mTESR（freeder-free medium）,and the classical ES culture medium were chosen for culture of iPS cells.The iPS cells maintaining in above mentioning culcure systems were co-cultured with 0P9 cells（murine bone marrow stromal cells） in vitro to be induced to differentiate into hematopoietic stem/progenitor cells.Flow cytometry and real-time quantitative PCR were used to detect the expression of specific hematopoietic markers and the effects of different culture systems on the differentiation of iPS in vitro.Result：iPS cultured in the 3 selected medium could be differentiated into hematopoietic stem cells.Efficiency of hematopoietic differentiation was up to 28.4%in classical ES culture system,which was significantly higher than that in E8 and mTESR system.Conclusion：Under the co-culture with 0P9,iPS can differentiate into hematopoietic stem/progenitor cells,which shows higher efficiency when iPS maintained in the ES medium.