成人脂肪间充质干细胞体外长期培养及成骨潜能的实验研究

发布时间：2018-01-19 00:29

本文关键词： 脂肪组织干细胞传代诱导分化骨组织生物学特性　出处：《兰州大学》2009年硕士论文　论文类型：学位论文

【摘要】： 目的:观察体外长期培养对成人脂肪间充质干细胞(ADMSCs)的基本生物学性状及成骨分化潜能的影响。方法:从成人脂肪组织分离培养ADMSCs,体外传代扩增,流式细胞术鉴定细胞表型,倒置相差显微镜及透视电子显微镜观察细胞形态及超微结构,MTT法绘制生长曲线,成骨诱导,碱性磷酸酶(ALP)染色、Von Kossa染色及免疫细胞化学染色行成骨鉴定,分析光密度值。结果:成人ADMSCs表达CD44,不表达CD34,体外长期培养其形态、生长方式及超微结构无明显变化,成骨诱导后变为椭圆形、岛状分布,ALP、钙化结节、骨钙素(OC)及Ⅰ型胶原染色均呈阳性,但随着传代次数的增加,光密度值有所减低。结论:成人ADMSCs经体外长期培养其一般生物学性状可保持稳定,但成骨分化潜能有所下降。
[Abstract]:Aim: to observe the effects of long-term culture on the basic biological characteristics and osteogenic differentiation potential of adult adipose mesenchymal stem cells (ADMSCs). Methods: ADMSCs were isolated and cultured from adult adipose tissue. The phenotypes of the cells were identified by flow cytometry. The morphology and ultrastructure of the cells were observed by inverted phase contrast microscope and fluoroscopic electron microscope. MTT method was used to draw the growth curve, osteogenesis induction, alkaline phosphatase (ALP) staining, Von Kossa staining and immunocytochemical staining for osteogenesis identification and optical density analysis. Results: adult ADMSCs expressed CD44, but not CD34. The morphology, growth pattern and ultrastructure of adult ADMSCs were not changed after osteogenesis induction. ALP, calcified nodules, osteocalcin (OC) and type I collagen staining were positive, but the optical density decreased with the increase of passage times. Conclusion: the general biological characteristics of adult ADMSCs cultured for a long time can be kept stable, but the osteogenic differentiation potential of adult ADMSCs can be decreased.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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