人骨髓间充质干细胞的分离及鉴定

［摘 要］ 目的：建立分离并鉴定人骨髓间充质干细胞（HMSC）的方法。方法：联合应用Percoll（1.073 g•L^-1）密度梯度离心和贴壁筛选的方法分离HMSC进行体外培养，利用免疫细胞化学、细胞化学和电镜等技术手段对体外培养的HMSC进行鉴定。结果：HMSC具有独特的表型特征,即CD29、CD44阳性和CD34、CD45阴性。细胞糖原反应（PAS）阳性、苏丹黑反应（SB）阴性。电镜观察结果提示细胞较幼稚。结论：利用密度梯度离心和贴壁筛选的方法可以分离HMSC，联合应用免疫细胞化学、细胞化学和电镜等技术手段鉴定HMSC的方法有效。

Isolation and identification of human mesenchymal stem cells

Objective To establish a method to isolate and identify human mensenchymal stem cells(HMSC). Methods The HMSC were isolated and cultured in vitro by applying methods of density-grad centrifugal and anchoring screening. The immunocytochemistry, cytochemistry, and electron microscopy were used to identify HMSC cultured in vitro. Results The HMSC have particular phenotype:CD29 and CD44 were positive,CD34 and CD45 were negative.PAS was positive,and SB was negative.HMSC were infantile under electron microscope. Conclusion The HMSC can be isolated with density-grad centrifugal and anchoring screening. The method to identify HMSC with immunocytochemistry, cytochemistry and electron microscopy is effective.