小鼠脂肪来源基质细胞向成骨细胞的分化

Differentiation of Adipose Derived Stromal Cells into Osteoblasts in Mouse

目的研究小鼠脂肪来源基质细胞的分离培养、鉴定及向成骨方向分化的潜能。方法取小鼠睾丸脂肪垫用胶原酶消化后获得血管基质层(SVFs),进行体外培养。取第4代细胞用流式细胞技术检测细胞表面分子CD29、CD44、Sca-1,用碱性磷酸酶染色及矿化结节染色法鉴定分化的成骨细胞。结果小鼠脂肪来源基质细胞高表达CD29、CD44,双阳性达84.9%,但Sca-1成阴性表达。经诱导的成骨细胞碱性磷酸酶染色胞质成黑色,矿化结节-茜素红与Von Kossa染色成红褐色与黑色。结论小鼠脂肪来源基质细胞为一种理想的成体干细胞,可用于基础实验研究,且能成功分化为成骨细胞。

Objective To study the isolation, culture, identification and osteogenic differentiation potential of mouse adipose derived stromal cells（ADSCs）. Methods Stromal vascular fraction cells（SVFs） were obtained from mouse testis fat pad with type I collagenase digestion. The cell surface molecules of the 4th passage cultured cells were detected by flow cytometry and their osteogenic differentiation was identified by alkaline phosphatase and mineralized nodule staining. Results Cell surface molecules CD29 and CD44 were highly expressed in mouse ADSCs respectively, with 84.9% double positive , but negative for Sca-1. The cytoplasm of the induced cells became black by alkaline phosphatase staining, the mineralized nodule became reddish-brown and black by Alizarin red and Von Kossa staining respectively. Conclusion Mouse adipose derived stromal cells are a ideal source of the adult stem cells and can be differentiated into osteoblasts and widely used in the research.