摘要
骨髓间充质干细胞来源有限,数量稀少,极大地限制了其在组织修复、细胞治疗等方面的应用。为了解决细胞数量限制的问题,从提高分离效率的角度对骨髓间充质干细胞的分离过程进行了研究,考察了培养基种类、初次换液时间、起始接种密度对兔骨髓间充质干细胞分离效率的影响。结果表明,α-MEM培养基较DMEM、DMEM-LG培养基更适于兔MSCs的分离;从骨髓中分离得到的单个核细胞以1×106 cells/ml的密度接种,接种24h后换液在短时间内得到的MSCs 数目最多。采用以上分离条件,能够获得形态均一、可稳定传代10代以上仍保持旺盛增殖能力的MSCs,并具有向骨、脂肪分化能力。
Mesenchymal stem cells(MSC) are an attractive target for use in cell and gene therapy. One of the biggest challenges facing the clinical application is to obtain enough cells. The isolation process was examined. The effects of medium kinds, medium exchange time and initial plating density on isolation of rabbit bone marrow mesenchymal stem cells was studied to optimize the isolation conditions and improve the yield of MSCs. It was showed that the characteristics of α-MEM was better than that of DMEM and DMEM-LG for MSCs colony formation. The optimal yield of MSCs was obtained when mononuclear cells were cultured at a plating density of 1×106 cells/ml, and the medium was exchanged to remove the nonadherent cells after 24h of inoculation. MSCs isolated under above conditions maintained a homogenous morphology, the osteogenic and adipogenic potential over 10 passages.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2005年第6期15-19,共5页
China Biotechnology
基金
国家"863"计划生物与现代农业技术领域组织器官工程重大专项(2003AA205120)
