髓核细胞共培养诱导骨髓间充质干细胞向类髓核细胞分化  

Differentiation of bone marrow mesenchymal stem cells into nucleus pulposus-like cells after co-culture with nucleus pulposus cells

作　　者：韦葛堇[1] 黄育强[1] 覃万安[1] 廖长川 林舟丹[1] 

机构地区：[1]解放军第303医院骨科,广西壮族自治区南宁市530021

出　　处：《中国组织工程研究》2013年第45期7834-7839,共6页Chinese Journal of Tissue Engineering Research

基　　金：广西自然科学基金项目(桂科自0991287)~~

摘　　要：背景:骨髓间充质干细胞近年来作为种子细胞被广泛应用于髓核组织工程。在诱导骨髓间充质干细胞向髓核细胞分化的过程中,需要提供适当的细胞外微环境以及生长因子。目的:研究体外与髓核细胞非接触共培养和生物诱导剂条件下,兔骨髓间充质干细胞向类髓核样软骨细胞分化的可行性。方法:用单层培养法分别分离培养兔骨髓间充质干细胞和髓核细胞。选用第3代的髓核细胞和骨髓间充质干细胞置于Transwell培养皿上下层构建共培养体系并加入转化生长因子β1为主的生物诱导剂诱导培养21 d,以高糖DMEM培养基单独培养的骨髓间充质干细胞作为阴性对照。结果与结论:分离培养的兔骨髓间充质干细胞细胞表面CD29和CD44表达阳性,CD34、CD45的细胞表达阴性。与髓核细胞共培养诱导21 d后,可观察到骨髓间充质干细胞形态向多角形类圆形转变,胞质中分泌Ⅱ型胶原颗粒明显增多,RT-PCR结果表明诱导后细胞中Ⅱ型胶原mRNA表达明显增高。结果说明骨髓间充质干细胞在与髓核细胞共培养加生物诱导剂条件下可成功诱导分化为类髓核细胞。BACKGROUND： Bone marrow mesenchymal stem cells in recent years have been widely used as seed cells for nucleus pulposus tissue engineering. Appropriate extracellular microenvironment and growth factors are required for the induction of bone marrow mesenchymal stem cells differentiating into nucleus pulposus cells. OBJECTWE： To observe the differentiation of rabbit bone marrow mesenchymal stem cells into nucleus pulposus-like cells in vitro induced by biological inducers and co-culture conditions. METHODS： Rabbit bone marrow mesenchymal stem cells and nucleus pulposus cells were isolated and cultured using monolayer culture method, respectively. Passage 3 bone marrow mesenchymal stem cells were placed in the Transwell culture plates for co-culture with the passage 3 nucleus pulposus cells and then were induced biologically to differentiate into the nucleus pulposus-like cells for 21 days. Bone marrow mesenchymal stem cells were cultured alone in the high-glucose Dulbecco＇s modified Eagle＇s medium as negative controls. RESULTS AND CONCLUSION： The expressions of CD29 and CD44 were positive and CD34 and CD45 were negative in the bone marrow mesenchymal stem cells. After 21 days of co-culture with nucleus pulposus cells, the morphology of bone marrow mesenchymal stem cells changed obviously from polygonal to oval shape, and collagen type II granules were increased significantly. Reverse transcription-PCR results showed that expression of collagen type II mRNA was significantly increased after induction. These findings indicate that bone marrow mesenchymal stem cells can be induced into differentiate into nucleus pulposus-like cells in co-culture plus biological inducer conditions in vitro.

关 键 词：干细胞 骨髓干细胞 骨髓间充质干细胞 细胞共培养 诱导分化 转化生长因子β1 Ⅱ型胶原 髓核细胞 类髓核细胞 省级基金 干细胞图片文章 

分 类 号：R394.2[医药卫生—医学遗传学]