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目的:研究左归丸含药血清对间充质干细胞骨向分化过程中碱性磷酸酶含量变化的影响。方法:采用全骨髓贴壁法培养间充质干细胞,连续进行3次差速贴壁法纯化间充质干细胞。常规诱导组以β-甘油磷酸钠、地塞米松、维生素C诱导间充质干细胞骨向分化,实验组在常规诱导组基础上加以左归丸含药血清。使用茜素红染色钙结节、碱性磷酸酶染色鉴定诱导成功。分别在0,7,14,21 d用比色法检测细胞中碱性磷酸酶含量。结果:全骨髓贴壁法和差速贴壁法联合使用可得到较为均一间充质干细胞。用比色法测得碱性磷酸酶含量随着诱导时间延长而增加。结论:大鼠间充质干细胞经药物诱导后可以分化为成骨细胞。左归丸含药血清能够促使诱导过程中碱性磷酸酶含量表达增加。
Objective: To study the influence of Zuogui pill containing serum on the change of alkaline phosphatase in the process of bone differentiation of mesenchymal stem cells. Methods: Mesenchymal stem cells were cultured by whole bone marrow adherent method and mesenchymal stem cells were purified by differential adherent method three times in a row. Conventional induction group induced osteogenic differentiation of mesenchymal stem cells with β-glycerophosphate, dexamethasone and vitamin C. The experimental group was given Zuogui pill serum on the basis of routine induction group. Alizarin red stained calcium nodules, alkaline phosphatase staining identified induction success. The alkaline phosphatase contents of the cells were detected by colorimetry at 0, 7, 14 and 21 days respectively. Results: The combination of whole bone marrow adherent method and differential adherent method can obtain more homogeneous mesenchymal stem cells. Alkaline phosphatase content increased with induction time by colorimetry. Conclusion: The rat mesenchymal stem cells can differentiate into osteoblasts after being induced by drugs. Zuogui pill containing serum can promote the induction of alkaline phosphatase expression increased.