摘要
目的体外培养骨髓单个核细胞（BMNCs），并观察其成脂肪细胞分化能力。方法无菌取SD大鼠骨髓制作细胞悬液，差速贴壁法培养第三次贴壁细胞，传代扩增，取3-5代细胞用于实验。流式细胞分析检测CD34、CD44、CD45、CD90、CD144（VE—cadherin）和血管内皮细胞生长因子受体．2（KDR）表达；STEMPRO Adipogenesis Differentiation Kit检测细胞成脂肪分化能力。结果通过差速贴壁法可获得相对单一的BMNCs，流式细胞分析表明培养的BMNCs细胞表达CD34、CD44、CD45及CD90等表面标志，但不表达CDl44和KDR；在未经诱导的情况下，培养的BMNCs能自发分化为脂肪细胞；培养BMNCs成脂肪细胞诱导后在第3、5、7d，成脂率分别为（22．4±3．8）％、（55．6±11．4）％以及（82．4±17．7）％，显著高于诱导培养第1d[（1．3±1．1）％，P〈0．011。结论骨髓单个核细胞体外培养后可以诱导分化为脂肪细胞。
Objective To observe the capacity of in vitro cuhured bone marrow mononuclear cells （BMNCs） to differentiate into fat cells, Methods The bone marrow of SD rats was sampled in a sterile manner to prepare cell suspension. The third adherent cells were cultivated by differential adhesion method for passage and amplification. Passage three （P3）-passage five （P5） of the third adherent cells were used in the experiments. The expressions of CD34, CD44, CD45, CD90, CD144 （VE-cadherin） and the vascular endothelial growth factor receptor-2 （KDR） were detected by flow cytometry, the capacity of cells to differentiate into fat cells was detected by STEMPRO Adipogenesis Differentiation Kit. Results Relatively homogeneous BMNCs were obtained by differential adhesion method. Flow cytometry indicated that the cultured BMNCs cells expressed CD34, CD44, CD45 and CD90 while did not express CD144 and KDR. Even in the absence of adipocyte differentiation induction, the cultured BMNCs might be spontaneously differentiated into fat cells. On the 3rd, 5th and 7th day since BMNCs were induced to fat cells, the fat cell percentages were （22.4± 3.8）%, （55.6±11.4）% and （82.4±17.7）% respectively, which were all significantly higher than that on the first day of induction culture [（1.3±1.1）%, P 〈 0.01]. Conclusion BMNCs cultivated in vitro may be differentiated into fat cells.
出处
《西南国防医药》
CAS
2016年第12期1382-1385,共4页
Medical Journal of National Defending Forces in Southwest China
基金
国家自然科学基金(81170316)
国家科技支撑计划项目(2014BI01B00)
云南省科技创新团队建设项目(20140810)
