人骨髓基质干细胞体外培养和生物学特性鉴定

Cultivation in vitro and identification of biological characteristics with human bone marrow stromal cells (hBMSCs)

目的体外分离、纯化培养人骨髓基质干细胞(hBMSCs),对其表面标志物、多向分化特性进行鉴定,为骨组织工程提供理想的种子细胞来源。方法抽取健康成人骨髓,采用全骨髓培养法,多次消化以纯化hBMSCs;应用流式细胞仪对细胞表面标志物进行检测;采用成骨、成软骨和成脂培养基进行三向诱导分化。结果培养的第3代细胞表面抗原检测显示:表达CD105、CD166和CD29,不表达造血和内皮标志CD34、CD45和CD14,在体外分别诱导出其向成骨、成软骨和成脂肪细胞分化。结论hMSCs体外经分离传代培养后细胞成分较为单一,具有多向分化潜能,符合间充质干细胞的基本功能特征。

[Objective] To establish a method for isolation and cultivation of adult hBMSCs in vitro and explore its biological characteristics for obtaining ideal seed ceils to be used in bone tissue engineering. [Methods] Bone marrow was obstracted from the healthy adult human and culture-expanded in vitro by the method of total marrow. The hBMSCs were purified by discarding suspended cells through exchanging medium . For identifying hBMSCs, cells surface antigens, including CD105, CD166, CD20, CD34, CD45 and CD14 were detected by flow eytometry. Then hBMSCs were induced with the special inducer, its trilineage differential potential into osteoblast, ehondroeyte and adipocyte were observed by using microseope.[Results] The result showed that the third passage hBMSCs highly expressed CD105, CD166 and CD39, and were negative for any hematopoietic and endothelial markers （CD34, CD45 and CD14）. Functionally, they could differentiate into the osteoblast, chondrocyte and adipoeyte by induced respectively with the special inducer. [Conclusion] The cultured cells had only composed of undifferentiated hBMSC and the potential of multilineage differentiation. They had the properties of stem cells.