体外诱导人骨髓间充质干细胞向造血细胞分化的研究

目的探讨体外诱导人骨髓间充质干细胞（hMSC）向造血细胞分化的可行性及分化条件。方法首先建立hMSC的分离培养扩增体系，制备小鼠胎肝基质细胞条件培养液（FLSC-CM），将hMSC接种在分别含FLSC-CM和IL-6、SCF组合的培养体系中，通过形态学、直接免疫荧光染色检测法、粒-单／巨噬系集落形成细胞培养（CFU-GM）对分化细胞进行鉴定。结果hMSC经FISC—CM诱导培养9天后，产生较多的悬浮细胞，悬浮细胞瑞士染色后形态类似于单核或小淋巴细胞，表达人CD34和人CD45表面抗原。且能够形成造血祖细胞集落（CFU-GM）。结论FLSC-CM可诱导hMSC分化为具有造血干／祖细胞特性的前体细胞。

Study on hematopietic differentiation of human mesenchymal stem cells derived from bone marrow in vitro

Objective To explore the feasibility and condition of the hematopoietic differentiation of hMSC in vitro.Methods hMSC derived from adult bone were isolated,culture-expanded efficiently in vitro.Fetal liver stromal cell conditioned(FLSC-CM) were well prepared Culture-expanded hMSC were cultured with FLSC-CM and the combination of IL-6 and SCF.the hematopoietic differentiated cells type of hMSC were characterized by morphology,CFU-GM assay,and immunofluorescence methods.Results There were lots of suspension cells derived from hMSC cultured with FLSC-CM after 10 d.Immunofluorescence analysis of the suspension cells demonstrated the presence of the positive CD34 and CD45 cells which were labeled by both PE and FITC,and appear red and green respectively.The suspension cells remained viable with the morphology similar to monocytes or small lymphcytes,and had the ability to form hematopoietic progenitor cells colonies in methylcellulose culture system.Conclusion The hMSC can be successfully directed toward their differentiation into progenitor cells with the characteristics of hematopoietic stem cells/hematopoietic progenitor cells after cultivation of FLSC-CM.