小鼠骨髓间充质干细胞的分离培养及生物性状研究

The Isolation,Culture and Study on Their Biological Features in vitro in Mouse Bone Marrow - derived Mesenchymal Stem Cells

目的:分离培养小鼠骨髓间充质干细胞,研究其体外培养的生物特性。方法:获取骨髓间充质干细胞进行培养,倒置相差显微镜观察细胞生长情况,绘制生长曲线,冷冻保存细胞复苏后的生长状况观察。免疫细胞化学法鉴定细胞表面抗原。在地塞米松、β-甘油磷酸钠、抗坏血酸的作用下,进行成骨诱导分化。结果:原代和传代培养的细胞体外培养细胞呈现梭形、多角形外观,具有较强的生长增殖能力,复苏细胞生长状况无明显改变;细胞CD44,CD54抗原有阳性表达,CD34呈阴性表达。分化细胞表现成骨细胞特性,合成碱性磷酸酶(ALP)能力增强,矿化结节逐渐出现。结论:建立稳定可靠的分离培养小鼠骨髓MSCs方法,分离出的MSCs具有干细胞的生物特性,可用于骨组织工程中的种子细胞。

Objective: To culture mesenchymal stem cells (MSCs) derived from mouse bone marrow, study on their biological features in vitro. Methods: Mouse MSCs was obtained and cultured,the morphology of cells and growth curves of MSCswere studied with Phase Contrast Microscope.The biological features of MSCs and Cryogentic storged cells studied.The cells surface antigens were employed to detect.Induced and differentiated MSCs into osteoblasts with dexamethasone,β-glicerophosplale,AsAP. Results: The cultured cells appeared morphologically as spindle - shaped and plygonal - shaped. MSCs showed active proliferative in vitro in primary and passage cultures. Cryogentic storaged cells biological features did not alterate. Cells were positive for CD44, cdS4 and negative for CD34. Differentiation cells showed the specific property of osteoblast, the ability of secreting ALP was enhanced, prmoted expression of mineralized nodular, Cryogentic storaged cell was so. Conclusion: A reliable method was established for obtain and culture mouse bone marrow MSCs in vitro, the bone marrow MSCs have biological features of stem cells and being used as the seed cells in the osteogentic tissue engineering.