小鼠骨髓间充质干细胞生物学特性和体外诱导分化

Murine marrow-derived mesenchymal stem cell: biological characteristics and induced differentiation in vitro

目的研究小鼠骨髓间充质干细胞的生物学性状和多系分化潜能。方法取Balb/c小鼠骨髓单个核细胞在低糖的培养液中培养出贴壁生长的细胞,进行形态学观察、细胞周期和免疫表型分析;在不同的因子作用下诱导向成骨细胞、软骨细胞,脂肪细胞分化,并检测诱导后细胞相应的基因表达。结果小鼠骨髓间充质干细胞贴壁生长后形态较均一,增殖能力随着传代逐渐增强,但从第8代后增殖能力明显减退。细胞表达CD2 9,CD38,CD4 4 ,CD10 6等标记,但CD34和H -2k表达阴性。在不同的诱导培养体系里间充质干细胞能分化为成骨细胞、软骨细胞和脂肪细胞,相应的骨钙蛋白基因,Ⅱ型胶原基因,脂蛋白脂酶基因表达都明显增强。结论从小鼠骨髓可以分离培养出间充质干细胞,在体外有效扩增和诱导分化。表明可以以小鼠为模型研究间充质干细胞在组织工程、细胞移植、基因治疗等领域的运用。

To explore the biological characteristics and in vitro differentiating potential of murine marrow-derived mesenchymal stem cells. Methods The techniques applied in isolation of murine marrow-derived mesenchymal stem cells were described, as well as the methods to directing their differentiation into osteogenic, chondrogenic, and adipogenic lineages. Their characterizations regarding morphology, surface marker profiles and growth kinetics have also been explored. Results These cells were positive for the surface markers CD29, CD38, CD44 and CD106, and exhibited a homogeneous, distinctive morphology, and were shown to differentiate along osteogenic, chondrogenic, and adipogenic pathways. After the 8th passage, MSC had a markedly diminished proliferation rate. Conclusion The methodology for the culture and induced differentiation of mouse MSCs, as well as a comprehensive understanding of their biology, will make the development of cellular and genetic therapy protocols in murine models possible, and provide new information in the field of adult stem cells research.