小鼠骨片间充质干细胞分离培养及鉴定  

Isolation,culture,and characterization of mouse bone derived mesenchymal stem cells

作　　者：马予洁[1] 陈津[1] 黄梁浒[1] 王庆华[1] 郭子宽[1] 谭建明[1] 

机构地区：[1]南京军区福州总医院全军器官移植研究所 福建省移植生物学重点实验室 福建省干细胞应用工程技术研究中心,福州350025

出　　处：《中华细胞与干细胞杂志（电子版）》2013年第4期6-9,共4页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：福建省科技重大专项(2012YZ0001);福建省科技创新平台建设项目(2010Y2006);军队十二五重点课题(BWS11J004);军队临床高新技术重大专项(2010gxjs026)

摘　　要：目的建立小鼠骨片间充质干细胞(MSC)分离培养及扩增的方法。方法取小鼠胫骨和股骨,洗去骨髓后,用胶原酶Ⅰ消化疏松骨密质,利用MSC具有迁徙和贴壁生长的能力进行分离。并对获取的细胞进行流式鉴定和诱导分化。结果培养2 d小鼠骨片边缘爬出成纤维样细胞,呈克隆和鱼群样生长,并可以进行持续传代培养。流式鉴定结果显示这群细胞表达MSC标志Scal1(92.7﹪),CD29(98.4﹪),CD90(91.6﹪),不表达造血细胞标志CD34(1.57﹪),CD45(3.99﹪),CD11b(0.63﹪),并可成功诱导分化成骨细胞和脂肪细胞。结论成功建立从小鼠骨片中获得MSC的方法,为实验研究提供可靠的细胞来源。Objective To establish a method for isolation and cultivation of mesenchymal stem cells from mouse bone. Methods Methods Tibia and femur were harvested from C57BL/6 mice, and bone marrow was flushed out of the cavity with ct-MEM medium. The bone was cut into fragments with scissors and digested with collagenase I for 1-2 hours. The digested bone fragments were cultured in the medium for mesenchymal stem cells. The harvested cells were characterized by flow cytometry and differentiation capacity. Results Fibroblast-like cells migrated out of the bone fragments, which grew in the medium clonally and was cultured for several passages. Flow cytometry showed that the cells expressed Scal-1 （92.7 %）, CD29 （98.4 %）, CD90 （91.6 %）, but negative for hematopoietic cells markers CD34 （1.57 %）, CD45 （3.99 %）, CDllb （0.63 %）. And cells could be induced to differentiate into osteoblast and adipocyte. Conclusion Mesenchymal stem cells were successfully isolated from mouse bone fragments.

关 键 词：小鼠 间充质干细胞 骨片 细胞培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]