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目的观察诱导时间的不同对大鼠脂肪间充质干细胞分化为成骨细胞的影响。方法脂肪间充质干细胞(ADSCs)分诱导组和非诱导组。诱导组在细胞接种同时(A组)、50%～60%融合时(B组)、85%～95%融合时(C组)分别加入同一浓度的由地塞米松、维生素C和β-甘油磷酸钠。非诱导组的A、B、C组由普通培养基培养不加入诱导剂。结果经钙钴染色后,诱导组大部分细胞呈阳性反应,以细胞达85%～95%融合时的结节体积最大。非诱导组未见片状阳性细胞。茜素红染色后可见红色结节,以细胞达85%～95%融合时诱导培养组形成结节较早。细胞内碱性磷酸酶活性检测表明诱导组与同期非诱导组相比细胞内碱性磷酸酶活性明显提高。结论 ADSCs经过诱导后,可向成骨细胞方向分化;诱导组细胞内碱性磷酸酶活性明显高于同期非诱导组;初始接种密度相同的情况下,ADSCs间相互接触程度越高,成骨能力越强。
Objective To observe the effect of different induction time on differentiation of rat adipose-derived mesenchymal stem cells into osteoblasts. Methods Adipose-derived mesenchymal stem cells (ADSCs) were divided into induction and non-induction groups. At the same time of inoculation (group A), 50% -60% of fusion (group B) and 85% -95% of fusion (group C), the same concentrations of dexamethasone, Sodium phosphate. The non-induced group A, B, C group by ordinary culture medium without adding inducer. Results After calcium-cobalt staining, most of the cells in the induction group were positive for the largest volume of nodules when cells reached 85% -95% confluency. Non-induced group no flaky positive cells. Alizarin red staining showed red nodules, with 85% ~ 95% of the cells fused to induce the formation of nodules earlier. Intracellular alkaline phosphatase activity test showed that the induction group compared with the same period non-induced group significantly increased intracellular alkaline phosphatase activity. Conclusion ADSCs can differentiate into osteoblasts after induction. The alkaline phosphatase activity in the induced group is significantly higher than that in the non-induced group at the same time. With the same initial inoculation density, the higher the degree of mutual contact between ADSCs, osteogenic capacity The stronger.