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目的 :分离、纯化小鼠卵黄囊间质干细胞 ,并观察其定向分化成骨、成软骨及成脂的潜能。方法 :分离、纯化孕 8.5d的小鼠卵黄囊间质干细胞 ,传代 4次后进行成骨、成软骨及成脂诱导。观察细胞形态 ,检测AKP ,Ⅰ ,Ⅱ ,Ⅲ型胶原及BMP - 1和BMP - 2的表达 ,并进行组化染色 ,检测成骨、成软骨及成脂情况。结果 :体外可获得纯化的卵黄囊间质干细胞 ,大小形态均一 ,呈梭形。成骨诱导后细胞向星形转化 ,AKP、Ⅰ型胶原、BMP - 2及VonKossa’s染色阳性 ,Ⅲ型胶原阴性。成软骨诱导后 ,细胞向多角形和圆形转化 ,AKP、Ⅱ型胶原、BMP - 1及Alcianblue染色阳性 ,Ⅰ型胶原阴性。成脂诱导后细胞肥大 ,油红O染色阳性。结论 :体外纯化的卵黄囊间质干细胞可诱导成骨、成软骨及成脂 ,具有多向分化潜能
OBJECTIVE: To isolate and purify mouse yolk sac mesenchymal stem cells and observe its potential for osteogenic, cartilage and adipogenic differentiation. Methods: The mouse yolk sac mesenchymal stem cells that had been pregnant for 8.5 days were isolated and purified. After being passaged for 4 times, osteogenic, cartilage and adipogenic induction were performed. The morphological changes of the cells were observed. The expressions of AKP, type Ⅰ, Ⅱ, Ⅲ collagen and BMP - 1 and BMP - 2 were detected, and the histochemical staining was performed to detect the osteogenic, cartilage and adipogenic status. Results: Purified yolk sac mesenchymal stem cells were obtained in vitro. After osteogenic induction, the cells transformed into a star with AKP, type Ⅰ collagen, BMP - 2 and VonKossa ’s stain, and type Ⅲ collagen negative. After chondrogenic induction, the cells transformed into polygons and circles, positive for AKP, type Ⅱ collagen, BMP - 1 and Alcianblue, type Ⅰ collagen was negative. After adipogenic induction, the cells were hypertrophy and positive with oil red O staining. Conclusion: The yolk sac mesenchymal stem cells purified in vitro can induce osteogenic, cartilage and adipogenic differentiation with multidirectional differentiation potential