大鼠脂肪间充质干细胞体外培养及其分化研究

目的:建立大鼠脂肪间充质干细胞体外分离培养体系并研究其多向分化潜能。方法:取6周龄大鼠双侧腹股沟和附睾处脂肪,酶解法分离培养原代脂肪间充质干细胞,绘制第三代细胞生长曲线。成脂诱导4 d,成骨诱导培养18 d,分别通过油红O染色、碱性磷酸酶活性检测和钙盐沉积VonKossa染色来观察细胞成脂、成骨表型转化。结果:大鼠脂肪中能分离培养出一定量的脂肪间充质干细胞,其群体倍增时间为(60±3.2)h。在成脂诱导培养液作用下可分化为脂肪细胞;成骨诱导培养液作用下,可特异性表达成骨分化标志碱性磷酸酶,同时也具有矿化细胞外基质的能力。结论:脂肪间充质干细胞具有易于取材、大量分离培养和定向诱导成骨分化的特点,是骨组织工程理想的种子细胞的来源。

Isolation culture and differentiation of the adipose derived adult stem cells from rats

Objective：To find out the optinial way of isolating and cul- turing the adipose derived adult stem cells from rats and investigate whether these cells would be induced differentially along osteogenie or adipogenic lineages in vitro. Methods ： Adipose derived adult stem cells were isoated from the fat tissues of rats aged 6 weeks and passage 3 cells were cultured in ostengenic or adipogenic media. Cells doubling time was calculated and the differentiation was detected by oil red O stain, alkaline pbosphatase assay or Von Kossa silver stain. Results ： At 18 days, cells differentiated into osteoblast in vitro demonstrated by alkaline phosphatase assay and bone nodule formation. At 4 days, cells differentiated into adipose in vitro demonstrated by oil red O stain. Cells doubling time was （ 60 ± 3.2 ） hours. Conclusion ： Adipose derived adult stem cells would be a promising source of adult autologous stem cells for bone tissue engineering.