小鼠密质骨间充质干细胞的分离、培养及鉴定

Isolation, culture and identification of mesenchymal stem cells from mouse compact bone

目的：建立一种易操作的、高效的小鼠间充质干细胞（mMSC）的分离培养方法，并对其表面标志和多向分化潜能进行鉴定，为mMSC的获取提供了新的实验依据。方法：选取2～3周龄C57BL／6小鼠，取出双侧股骨、胫骨，冲出其内骨髓细胞并弃掉，将骨段剪成小块，胶原酶消化后用组织块培养法培养出MSC。倒置显微镜观察其生长过程，流式细胞术检测其表面标志，并对其进行成脂及成骨鉴定。结果：使用该方法分离培养的mMSC形态均一、生长良好，细胞形态呈成纤维细胞样，表达CD29和Scal-1，不表达CD34、CD45和CDllb，并具有成骨成脂潜能。结论：利用本操作方法，可以从小鼠密质骨中成功分离培养得到MSC，且高效易行。

Objective： To establish an efficient method for isolation and culture of mesenchymal stem cells （MSC） from mouse compact bone, to identify surface markers of the cells and to provide experiment data for its application. Methods： 2-3 week mice were selected, firstly their bone marrow cells from the femur and tibia of both sides were rushed and discarded, and then these femur and tibia were dissected into small pieces which were subsequently digested by collagenase for further tissue culture to obtain MSC. The MSC were observed under a microscope, their surface markers were identified with FACS, and then their differentiation capacity into osteogenic and adipogenic cells were investigated. Results： The MSC isolated from mouse compact bone grew well and was homogeneous. The cells showed positive with CD29 and scal-1, but showed negative with CDllb~ CIM5 and CD34. They could differentiate into osteogenic and adipogenic cells. Conclusion： The method described here was easy and efficient, by which we could isolate and culture MSC from mouse compact bone successfully.