贵州医科大学学报

2013, (01) 5-9+15

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人脐带间充质干细胞的分离培养及超微结构特点研究
Isolation and Culture of Human Umbilical Cord Mesenchymal Stem Cells and Analysis on Their Ultrastructural Characteristics

张芬熙;洪艳;梁文妹;
ZHANG Fenxi,HONG Yan,LIANG Wenmei(Department of Histology and Embryology,Guiyang Medical College,Guiyang 550004,Guizhou,China)

摘要(Abstract):

目的:分离和培养人脐带间充质干细胞(hUCMSCs),研究其生物学特性及超微结构特点。方法:采用组织块植入法培养原代hUCMSCs,绘制细胞生长曲线,根据细胞生长状态选第3代hUCMSCs进行实验。流式细胞术、透射电镜和免疫细胞化学等技术研究hUCMSCs的生物学特性。结果:第1~2天为hUMSCs生长潜伏期,第3~5天为对数增长期,第6天进入平台期。hUCMSCs阳性表达CD44、CD90、CD29和CD105,阴性表达CD34和CD45。诱导培养可促使hUMSCs向成骨及成脂细胞分化。电镜下hUCMSC形态不规则,核大,核质比大,核仁明显,核内染色质稀疏,胞质较少,符合原始细胞的超微结构特点。结论:采用组织块植入法获得的hUCMSCs具有较强的增殖能力,在体外诱导培养下可向成骨和成脂细胞分化。
Objective: To isolate and culture human umbilical cord mesenchymal stem cells(hUCMSCs),and to study their biological feature and ultrastructural characteristics.Methods: The primary hUCMSCs were cultured by using tissue block implantation method.Cell growth curve was drawn.Based on cell growth condition,the 3rd passage cells were used in the experiment.Biological characteristics of hUCMSCs were analyzed by using flow cytometry,transmission electron microscope and immunocytochemistry.Results: The latent growth phase and logarithmic growth phase of hUCMSCs were in day 1 to day 2,and day 3 to day 5 respectively;After the 6th day,cell growth came into platform phase.In hUCMSCs,CD44,CD29,CD90 and CD105 were positively expressed,while CD34 and CD45 were negatively expressed.After inductive culture,hUCMSCs could differentiate into osteoblastic cells or adipogenic cells.Under transmission electron microscope,irregular cell shape,big cell nucleus,big karyoplasmic ratio and obvious nucleolus of hUCMSCs were observed.The chromatin sparsely distributed in the nucleus,and cytoplasm was in small quantity.All of these accorded with the ultrastructural characteristics of primitive cells.Conclusions: hUCMSCs obtained by tissue block implantation method have strong proliferation ability,and can be induced to differentiate into osteoblasts and lipoblasts in vitro.

关键词(KeyWords): 人;脐带;间充质干细胞;细胞周期;超微结构
persons;umbilical cord;mesenchymal stem cells;cell cycle;ultrasructure

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基金项目(Foundation): 国家自然科学基金(No:8110099);; 贵州省社会发展科技攻关项目,黔科合SY[2009]3071;; 贵州省教育厅项目,黔教高发[2010]305

作者(Author): 张芬熙;洪艳;梁文妹;
ZHANG Fenxi,HONG Yan,LIANG Wenmei(Department of Histology and Embryology,Guiyang Medical College,Guiyang 550004,Guizhou,China)

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