人骨髓基质干细胞克隆对脐血造血干细胞体外扩增作用的研究

目的：探讨人骨髓基质干细胞的克隆化培养及其对体外造血干细胞扩增的影响。方法：取正常肋骨分离骨髓，制备单个核细胞贴壁培养，约5d后取集落样生长的梭形细胞扩增培养，传代、将分选的脐血造血干／祖细胞接种到克隆培养的人骨髓基质干细胞及其他培养条件液上，比较不同培养条件及不同代次骨髓基质干细胞对造血干／祖细胞扩增能力及集落形成能力的影响。结果：人骨髓基质干细胞能扩增达20代以上，经流式细胞仪检测证实基质干细胞，单纯细胞因子组和细胞因子加基质干细胞组能有效扩增造血细胞，而后者有维持长期造血的作用。结论：该克隆培养方法能有效扩增骨髓基质干细胞，培养的细胞有体外支持长期造血的作用。

A STUDY ON THE CLONE CULTURE OF HUMAN BONE MARROW MESENCHYMAL STEM CELLS (hMSCs) AND THEIR EX VIVO SUPPORT AFFECT ON HEMATOPOIESIS

Objective:To study the clone culture method of hMSCs and their hematopoiesis support affect.Methods:Clone culture the hMSCs, put the positive magnetic sorted umbilical cord blood CD34+ hematopoietic stem/progenitor cells (HSC) under different conditional culture media with or without hMSC feederlayer referred to P5 and P15 of clone cultured hMSCs respectively, measure the enlargement fold of the proliferated cells and the CFCs cultured on colony-forming-unit culture condition.Results:The clone cultured hMSCs detected by FACS can be passaged for up to 20 passages the CD34+ HSC cells can proliferate effectly trader the culture conditions of hematopoiesis growth factors alone or hematopoiesis growth factors with hMSC layer ,the CFCs formed on hMSC layer of different passages are more than that of the group without a hMSC layer for long term culture. The P5 MSC possesses a better hematopoietic surport affect than the P15.Conclusion: The hMSCs mn proliferate ex vivo effectly under the culture condition, they can support long term hematopoiesis.