成人骨髓间充质干细胞体外诱导分化为软骨细胞的实验研究

Expermental Study on Differentiation of Human Bone Marrow-derived Mesenchymal Stem Cells into Chondrocytes in Vitro

目的研究体外三维培养条件下,成人骨髓间充质干细胞(BMSCs)定向分化软骨细胞可行性。方法用密度梯度离心法分离纯化BMSCs,并传代扩增后,用离心三维培养法在软骨诱导剂下进行诱导培养,以常规培养液培养的BMSCs作为阴性对照,于诱导培养第21天取出标本行苏木精-伊红染色、甲苯胺蓝染色、s-100蛋白免疫组织化学检测。结果BMSCs经21d的离心三维诱导培养后,培养管出现软骨外观组织块,组织切片甲苯胺蓝染色呈明显异染,s-100蛋白免疫细胞化学检测阳性。对照组阴性。结论成人BMSCs在三维培养条件下可成功诱导分化为软骨细胞。

Objective To study the feasibility of inducing adult bone marrow mesenchymal stem cells into ehondrocytes in three - dimensional condition. Methods Cells obtained in bone marrow aspirates by density gradient centrifugation were cultured in monolayer from adult and then transferred into tubes and allowed to form three- dimensional aggregates in a chemically defined medium including TGF - β1 insulin dexamethasone etc. BMSCs formed the aggregates in the conventional culture medium as negative control. These two aggregates were harvested after 21 - day＇s chondrogenic induction. For histological evaluation, sections were stained with haematoxylineosin and toluidine blue. For immunohistochemistry evaluation, sections were incubated with the monoclonal antibody specific to s- 100 protein. Results BMSCs were cultured in three- dimensional condition for 21 days with defined medium and were metachromasia by toluidine blue and could be identified by positive staining for s- 100 protein. The control group was not. Conclusion Adult BMSCs can be induced into chondrocytes in three - dimensional condition by the chemically defined medium.