摘要
为探索大鼠骨髓间充质干细胞的最佳培养条件及表型,用大鼠股骨抽取骨髓,密度梯度离心分离骨髓间充质干细胞。通过测定生长曲线和克隆形成能力,比较不同培养条件、不同融合度传代及不同生长基质对MSC的影响。免疫组化双标法检测增殖MSC的表型。结果Matrigel包被培养器皿,含10％胎牛血清、10 ng/mL EGF的α-MEM培养基,50％融合时消化传代,是MSC保持低分化状态、体外大量扩增的较佳条件。绝大部分MSC表达CD44,部分MSC表达c-kit,MSC几乎不表达CD34。说明以上培养条件是MSC保持低分化状态又大量增殖的较好培养条件,为MSC用于细胞移植和组织工程打下了良好基础。
To explore the best cultural conditions for rat bone marrow mesenchymal stem cells and to ientify its phenotype, bone marrow was aspirated from the femurs, then MSCs were obtained with density gradient centrifugation. The growth curve and clone forming efficiency were evaluated when MSCs were cultured in different medium, subcultured at different confluence, and plated on several substratum. Phenotype of MSCs was analyzed by immunohistochemical technique. When plated on matrigel, cultured at α-MEM (added 10% FCS, 10 ng/mL EGF), MSCs propagated rapidly and retained plouripotency. Most of MSCs were positive stained by CD44, some positive by c-kit, almost all negative by CD34. It is conclued that MSCs show easy survival in the subculture and rapid proliferation in present culture condition. MSCs will be excellent seed cells for the future cell transplantation.
出处
《科学技术与工程》
2003年第6期547-550,共4页
Science Technology and Engineering
基金
国家自然科学基金(30271663)
