小鼠骨片间充质干细胞分离培养及鉴定

目的建立小鼠骨片间充质干细胞（MSC）分离培养及扩增的方法。方法取小鼠胫骨和股骨，洗去骨髓后，用胶原酶I消化疏松骨密质，利用MSC具有迁徙和贴壁生长的能力进行分离。并对获取的细胞进行流式鉴定和诱导分化。结果培养2d小鼠骨片边缘爬出成纤维样细胞，呈克隆和鱼群样生长，并可以进行持续传代培养。流式鉴定结果显示这群细胞表达MSC标志Scall（92．7％），CD29（98．4％），CD90（91．6％），不表达造血细胞标志CD34（1．57％），CD45（3．99％），CD11b（0．63％），并可成功诱导分化成骨细胞和脂肪细胞。结论成功建立从小鼠骨片中获得MSC的方法，为实验研究提供可靠的细晌实源．

Isolation,culture, and characterization of mouse bone derived mesenchymal stem cells

Objective To establish a method for isolation and cultivation of mesenchymal stem cells from mouse bone. Methods Methods Tibia and femur were harvested from C57BL/6 mice, and bone marrow was flushed out of the cavity with ct-MEM medium. The bone was cut into fragments with scissors and digested with collagenase I for 1-2 hours. The digested bone fragments were cultured in the medium for mesenchymal stem cells. The harvested cells were characterized by flow cytometry and differentiation capacity. Results Fibroblast-like cells migrated out of the bone fragments, which grew in the medium clonally and was cultured for several passages. Flow cytometry showed that the cells expressed Scal-1 （92.7 %）, CD29 （98.4 %）, CD90 （91.6 %）, but negative for hematopoietic cells markers CD34 （1.57 %）, CD45 （3.99 %）, CDllb （0.63 %）. And cells could be induced to differentiate into osteoblast and adipocyte. Conclusion Mesenchymal stem cells were successfully isolated from mouse bone fragments.