大鼠骨髓间充质干细胞分离培养及鉴定的相关研究  

The study of cultivating and identificating the rat bone marrow derived mesenchymal stem cells

作　　者：宋珂[1] 石琦[1] 青莹 曹颖光[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院口腔科,湖北武汉430030

出　　处：《临床口腔医学杂志》2015年第9期531-536,共6页Journal of Clinical Stomatology

基　　金：国家自然科学基金(81400490);湖北省自然科学基金(2014CFB388)

摘　　要：目的:原代培养并鉴定大鼠骨髓间充质干细胞作为骨组织工程的种子细胞。方法:取大鼠骨髓,贴壁培养法培养。绘制生长曲线,检测细胞周期。real time PCR检测不同代数BMSCs胞内成骨相关因子mRNA水平。流式细胞仪检测细胞表面标志物。并行成脂及成骨诱导。结果:原代取材后24 h细胞即可贴壁,7 d可生长达培养皿底面积的80%。P3代BMSCs的生长曲线呈S形,近90%的细胞处于G0/G1期。从P0到P5BMSCs中几乎都有内源性bFGF、Shh、Cbfa1、ALP、OC的表达。流式细胞仪检测99.7%的BMSCs胞膜表面同时表达CD29及CD90,不表达CD45。并具有成骨分化及成脂分化的能力。结论:原代培养的BMSCs为骨组织工程提供了可靠的细胞来源。Objective： The aim of this study was to cultivate and determine the bone marrow derived mesenchymal stem cells （BMSCs）. Method： Bone marrow harvested from rat femurs were cultured. BMSCs were selected and expanded through passaging in vitro. Furthermore, the biological properties of BMSCs were investigated by flow cytometry. Meanwhile the muhipotent differentiation characteristics were demonstrated using standard in vitro conditions. Result： Monoptychial heterogeneous cells were obtained. A total of 98.4 % of cells at passage 3 expressed cluster of differentiation （CD） 29 and CD90, but not CD45. The cells were able to differentiate into osteogenic and adipogenic ceils. Conclusion： BMSCs that are isolated from the rat bone marrow and exhibit the identified characteristics may be used as seed cells in bone tissue engineering.

关 键 词：骨髓间充质干细胞 骨组织工程 生物学性状 

分 类 号：Q813.1[生物学—生物工程]