不同年龄人骨髓间充质干细胞体外生长特性分析  

Study on in vitro growth characteristics of human bone marrow mesenchymal stem cells in different age adults

作　　者：潘兴华[1] 庞荣清[1] 张步振[1] 王鹤桦[1] 杨勇琴[1] 梁虹[1] 

机构地区：[1]成都军区昆明总医院医学实验科,云南昆明650032

出　　处：《生物医学工程与临床》2006年第5期275-279,共5页Biomedical Engineering and Clinical Medicine

基　　金：云南省自然科学基金重点项目资助(2003C013Z)

摘　　要：目的建立人骨髓间充质干细胞(marrow mesenchymal stem cells,MMSCs)体外分离培养和扩增方法,观察不同年龄人MMSCs在体外培养的生长特性,为MMSCs的临床应用提供技术方法和依据。方法采集10～60岁之间的人骨髓血,密度梯度法分离出单个核细胞,利用DMEM/F12培养基添加胎牛血清进行黏附培养和扩增MMSCs,分析第3代MMSCs生长情况和形态变化,细胞计数,用MTT法观察其增殖及生长特征,免疫组化法分析MMSCs的纯度。结果10～60岁人MMSCs均贴壁生长,呈典型MMSCs形态和生长特征,传代细胞的增殖速度比原代细胞快,但随着年龄的增长,原代及传代培养MMSCs的增殖能力下降,生长周期延长,其中CD105阳性细胞均大于95%,但老年人的CD105阳性细胞比率有所降低。结论密度梯度离心结合贴壁培养法可获得高纯度MMSCs,成人MMSCs在传代培养的增殖活性随年龄增加而降低。Objective To establish the in vitro isolation, culture and amplification method of human bone marrow mesenchymal stem cells （MMSCs）, and observe the in vitro growth characteristics of MMSCs in different ages. Methods An amount of 10 ml bone marrow was obtained from the iliac crest of healthy voluntear donors in different ages. The MMSCs were isolated by centrifuge and culutured in vitro. The proliferation and growth characteristics including morphological changes and cell count were studied by MTT essay in primary and passage culture. The purity of MMSCs was analyzed by immunohistochemical method. Results All the MMSCs obtained from 10～60 years adults grew into typical MMSCs morphology and growth characteristics. The proliferation rate of passage cells was higher than that of primary cells, but the growth and proliferation activity reduced while aging. Conclusion High purity MMSCs can be obtained by gradient centrifuging and adherent culture. The proliferation activity in passage cell culture of adult MMSCs is reducing while age increasing.

关 键 词：骨髓间充质干细胞 体外培养 年龄 生长特性 

分 类 号：Q813.11[生物学—生物工程]