ICR小鼠骨髓间充质干细胞分离及生物学特性的探讨
投稿时间:2015-05-26  修订日期:2015-07-08  点此下载全文
引用本文:杨超,辛娜,康炜,卢韬,董学君.ICR小鼠骨髓间充质干细胞分离及生物学特性的探讨[J].医学研究杂志,2016,45(1):96-99
DOI: 10.11969/j.issn.1673-548X.2016.01.024
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作者单位E-mail
杨超 710077 西安医学院第一附属医院检验科 superyang0210@163.com 
辛娜 710077 西安医学院第一附属医院检验科  
康炜 710077 西安医学院第一附属医院检验科  
卢韬 312000 绍兴市人民医院临床检验中心  
董学君 312000 绍兴市人民医院临床检验中心  
基金项目:陕西省教育厅专项科研基金资助项目(12JK0764);西安医学院第一附属医院科研基金资助项目(XYFY11-08)
中文摘要:目的 探讨分离、培养的ICR(Institute of Cancer Research,ICR)小鼠骨髓间充质干细胞(mouse bone marrow mesenchymal stem cells,mBM-MSCs)的细胞生物学特性。方法 取6~8周龄ICR小鼠,利用全骨髓反复贴壁和有限稀释培养法分离纯化mBM-MSCs,观察形态特点,测定生长曲线和活力,利用流式细胞仪分析细胞的周期和鉴定表面抗原,诱导其向成骨、软骨及脂肪细胞分化,采用染色法鉴定。结果 新分离的mBM-MSCs多呈小圆形,形态规整。培养传代后,细胞多变为梭形,大小较均匀,形态较一致。随着传代的次数增加,细胞的生长曲线、活力及周期呈现快速发育期、平台期和缓慢期;流式结果细胞CD44、CD73、SCA-1呈阳性反应,部分细胞CD90、CD105、STRO-1呈阳性反应,CD11b和CD45呈阴性反应;诱导分化为成骨细胞后其碱性磷酸酶、茜素红和Von Kossa银染色均呈阳性反应,分化成脂肪细胞后油红O染色呈阳性反应,分化成软骨细胞后阿尔新蓝染色呈阳性反应。结论 ICR小鼠骨髓中分离培养出的MSCs,生物学特点鲜明,适于做进一步研究。
中文关键词:ICR小鼠  骨髓间充质干细胞  细胞培养  分化  鉴定
 
Investigation on the Isolation and Biological Characteristics of Bone Marrow Mesenchymal Stem Cells of ICR Mice
Abstract:Objective To investigate the cell biological properties of isolated and cultured mBM-MSCs of ICR. Methods Mouse bone marrow mesenchymal stem cells of 6-8 weeks old ICR mice were in vitro isolated and purified by whole bone marrow repeated adherent culture and limiting dilution culture method. Cell morphology was observed. Its growth curve and vitality were measured, and its cell cycle and surface antigen markers were analyzed by flow cytometry and differentiation ability was detected to identify whether they are the BM-MSCs. Results The newly isolated mBM-MSCs were small round, regular shape. After subculture, cells size was uniform, and their morphology was consistent and multi-spindle. Along with the increased number of cell passage, the growth curve, viability and cycle of cells were showed the first rapid development stage, platform stage and slow activity period. Flow cytometry results showed that cell's surface maker CD44, CD73 and SCA-1 were positive, part of the cells CD90, CD105 and STRO-1 were positive, CD11b, CD45 were negative. After osteogenic, adipogenic and chondrogenic differentiation, alkaline phosphatase, alizarin red, Von Kossa silver, oil red O and alcian blue staining were positive respectively. Conclusion The biological characteristic of isolated and cultured mBM-MSCs of ICR was homogeneous and distinct. These cells were suitable for further research.
keywords:ICR mouse  BM-MSC  Cell culture  Differentiation  Identification
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