犬骨髓基质千细胞向软骨样细胞诱导分化的条件研究

目的研究犬骨髓基质干细胞在体外培养中定向诱导分化为软骨样细胞的最佳条件。方法从幼年犬骨髓中分离基质干细胞进行培养扩增，观察其生长特性，体外应用碱性成纤维细胞生长因子（b-FGF）和转化生长因子β1（TGF．B1）对第3代传代细胞诱导分化，并通过细胞的染色和Ⅱ型胶原免疫组化鉴定诱导分化细胞的类型。结果原代培养的基质干细胞集落融合周期约6～9d左右。第3代传代细胞经碱性成纤维细胞生长因子（b—FGF）和转化生长因子（TGF．131）诱导后，传代细胞在形态上呈软骨样改变；甲苯氨蓝异染性、阿新蓝染色阳性；Ⅱ型胶原免疫组化染色呈阳性。结论犬骨髓基质干细胞在体外培养在碱性成纤维细胞生长因子（b—FGF）和转化生长因子（TGF—β1）作用下可被诱导分化为软骨样细胞。

Study on Conditions of Canine Bone Marrow Stem Cells Differentiate into Chondrocytes

Objective To explore the best conditions of inducing bone marrow stem cells （BMSCs） to differentiate into ehondro- eytes in vitro. Methods BMSCs were isolated from juvenile canine by means of gradient centrifugation, purified by anchoring culture in vitro, and observed for their growth characteristics. After the administration of b-FGF and TGF-β1 with third generation passage cell,the induced differentiation of BMSCs was evaluated by staining and collagen Ⅱ immunocytochemistry. Results Af- ter 6 - 9 d, the fusion periods of colonies of BMSCs were achieved in vitro. After third generation, the passage cell differentiation were induced with b-FGF and TGF-β1 ,the cells became chondrocytes in morphology ;The cells were positive by Alcian blue stai- ning,toluidine blue staining and collagen Ⅱ immunoeytochemistry staining. Conclusion The BMSCs can be cultured and puri- fied well in vitre,and can also be induced to differentiate into chondrocytes-like cells by b-FGF and TGF-β1.