大鼠ADSCs的生物学特性及向成骨细胞分化的实验研究

目的探讨脂肪源性干细胞(ADSCs)的生长增殖活性以及成骨分化能力。方法取SD大鼠,无菌条件下取腹股沟部脂肪组织,制成单细胞悬液进行常规培养。倒置显微镜观察细胞生长情况,取第3代ADSCs,采用流式细胞技术检测细胞的增殖周期情况,茜素红染色观察诱导成骨情况。结果在倒置显微镜下所观察到的传代培养的ADSCs呈现长梭型及多角形的贴壁分布状态,形态大小比较均一,突起较少,并且增殖迅速,一般在3~4d即可生长汇合成单层、平行或者漩涡状排列。流式细胞仪检测细胞阳性率CD29为92.47%,CD44为90.05%,CD34为0.95%。成骨诱导21d,细胞钙结节形成明显,经茜素红染色出现红色结节的阳性染色,不加诱导剂的对照组染色为阴性。结论分离黏附细胞具有间充质干细胞的生物学特性,可以向成骨细胞分化。

The biological characteristics of adipose derived stromal cells and their differentiation into osteoblast in SD rat

Objective To investigate the biological characteristics of rat adipose derived stromal cells（ADSCs） and their differentiation into osteoblast in SD rat. Methods ADSCs were isolated from SD rat by density gradient centrifugation and cultured in vitro. The 3rd generation cells were obtained and cell generation cycle was examined by flow cytometry, and the osteogenis differentiation was analyzed by Von-Kossa staining. Results The ADSCs were found to be adherent to the wall and rapid proliferation with long spindle or polygonal or uniform shape with less processes, merged into a single layer parallel or swirly arranged 3-4d after culture. Flow cytomet13, showed that the positive rates were 92.47% for CD29, 90.05% for CD44, 0.95% for CD34 respectively. The positive hydroxyapatite nodules were found by Von-Kossa staining 21 days after culture. Conclusion ADSCs have the biological property of mesenchymal stem ceils and could be differentiated into osteoblast.