骨髓间充质干细胞体外培养和成骨诱导

目的：观察骨髓问充质干细胞（MSC）体外培养及诱导成骨分化的特征。方法：采用密度梯度离心和体外扩增培养人骨髓MSC，并进行成骨诱导。应用茜素红染色法和钙钴法分别进行成骨诱导的MSC钙化结节检测和碱性磷酸酶（ALP）检测。结果：MSC成骨细胞诱导培养第l～3天的细胞增殖旺盛，3～5d即融合成单层。随着诱导培养时间的延长，细胞逐渐堆积并矿盐沉积，形成矿化结节。培养14d和21d后，矿化结节形成率分别为（70．5±3．5）％和（87．5±3．5）%，ALP染色阳性率分别为（41．5±1．5）％和（85．2±1．7）％。结论：在体外培养条件下，骨髓MSC可诱导分化为具有含矿化结节及ALP阳性特征的骨样细胞。

Osteogenic differentiation of bone marrow mesenchymal stem cells during in vitro culture

Objective： To observe the characteristic of in vitro culture and osteogenic differentiation of bone marrow mesenchymal stem cells （MSC）. Methods： The human bone marrow MSCs were acquired through density gradient centrifugation, then expanded and cultured in vitro, and induced to differentiate into osteoblasts. The calcified nodules were determined by alizarin red staining （ARS）, and the alkaline phosphatase （ALP） was detected by the calcium cobalt method. Results： The bone marrow MSCs actively proliferated from day 1 to day 3, fused into monolayer cells after culture of 3 to 5 days. Then the gradual accumulation of cells and the precipitation of mineral salt formed mineral node with the extending culture and induction time. After incubation for 14 and 21 days, the formation ratesof mineral nodes reached （70.5±3.5）% and （87.5±3.5）% respectively, and the positive rates of ALP staining were （41.5±1.5）% and （85.2±1.7） % respectively. Conclusion： Under the in vitro culture condition, the bone marrow MSCs can be induced to differentiate into the ALP positive osteoblasts containing mineral nodes.