大鼠骨髓间充质干细胞分泌多种造血相关因子的研究

摘要

目的:初步探讨大鼠骨髓间充质干细胞(MSC)分离培养的方法及检测其所表达的多种细胞因子.方法:采用全骨髓贴壁法和单克隆培养法分离培养大鼠骨髓MSC,间接免疫荧光法鉴定其生物学特性,RT-PCR方法检测多种细胞因子的表达.结果:成功分离出大鼠骨髓MSC,CD44表达率为92.3%,CD45表达率为6.9%;RT-PCR分析示有干细胞因子(SCF)、血小板生成素(TPO)、粒细胞巨噬细胞集落刺激因子(GM-CSF)、转化生长因子(TGF-β1)的表达,不表达白介素-3(IL-3).结论:成功从大鼠骨髓中分离培养出MSC,并分泌多种造血相关因子,这可能揭示了其促造血功能的部分分子基础.%Objective : To study the rat bone marrow mesenchymal stem cells ( MSC ) isolation and culture methods and detect its expression in a variety of cytokines. Methods : Bone marrow adherent was cultured and isolated monoclonal bone marrow MSC, and identified hy indirect immunofluorescence biological characteristics. RT - PCR was used to detect the expression of multiple cytokines. Results: MSC was successfully isolated from rat bone marrow, CD44 expression was 92. 3％ , CD45 expression rate was 6. 9％; RT - PCR analysis showed that a stem cell factor ( SCF ), thrombopoietin ( TPO ) , granulocyte macrophage colony stimulating factor ( GM - CSF ) , transforming growth factor ( TGF - β1, ) expression, no expression of IL -3 ( IL - 3 ). Conclusion: MSC was successfully isolated from cultured rat bone marrow, and which secrete a variety of blood - related factors, which may reveal some of their hematopoietic function.