TGF-β3诱导骨髓间充质干细胞表达软骨细胞表型的实验研究

THE EXPERIMENT STUDY OF THE DIFFERENTIATION OF BONE MARROW MESENCHMAL STEM CELLS TO EXPRESS THE PHENOTYPE OF CHONDROCYTE LINEAGEBY TRANSFORMING GROWTH FACTOR-β3 IN VITRO

目的探讨TGF-β3体外诱导骨髓间充质干细胞分化为软骨细胞的可行性。方法分离培养骨髓间充质干细胞,鉴定其表面抗原,用含10%胎牛血清以及10ng/mL转化生长因子TGF-β3的条件培养基诱导,诱导后细胞通过软骨细胞特征性染色鉴定。结果骨髓间充质干细胞表面抗原CD29、CD44、CD105阳性,CD34、CD45、CD106以及HLA-DR阴性。诱导后细胞形态明显改变,甲苯氨蓝以及Ⅱ型胶原染色结果阳性。结论TGF-β3具有促进骨髓间充质干细胞在体外分化为软骨细胞的能力,可以作为组织工程种子细胞的一种有效来源。

Objective To investigate the feasibility of bone marrow mesenchymal stem cells into chondrocyte lineage by transforming growth factor-β3 in vitro. Method cells were isolated from bone marrow, and then were characterized by flow cytometry with a panel of monoelonal antibodies. Differentiation into chondrocyte-like cells was induced by cultivation of confluent cells in the presence of 10 % fetal calf serum and 10 ng/mL TGF-β3. These cells were characterized by immunohistochemistry stain. Result Isolated MSCs were positive for the markers CD105, CD29 and CD44 and negative for typical hematopoietic and endothelial markers CD34, CD45, CD106 and HLADR. The morphology of induced cells was obviously changed. Immunohistochemistry stain analysis of the confluent cells in situ showed positive for collogen type Ⅱ. Conclusion TGF-β3 has the ability to promote bone marrow mesenchymal stem cells differentiation chondrocyte lineage in vitro. The bone marrow MSCs may prove to be a useful source of tissue engineering seed cells.