骨髓单个核细胞的提取、分离、标记和体外培养

目的提取分离骨髓单个核细胞（BM—MNCs）。方法无菌条件下行髂后上嵴穿刺抽取小型猪骨髓20-40ml，采用Ficoll密度梯度离心法分离BM—MNCs。结果经2％台盼蓝鉴定细胞存活率为96％～99％，DAPI染色标记率100％，标记的细胞核发蓝色荧光。BM—MNCs的体外培养可见细胞呈梭形生长，通过CD44免疫组化鉴定证实为CD44＋骨髓间充质干细胞（MSCs）。结论BM—MNCs易分离，操作简便，可作为移植细胞选择。

Isolation, labeling and culture of bone marrow-derived mononuclear cells

Objective To isolate bone marrow-derived mononuclear cells （BM-MNCs）. Methods To aspirate 20-40 ml of marrow blood from the posterior iliac crest of swines under sterilization. Mononuclear cells were isolated by Ficoll density gradient centrifugation. Results Dyeing with trypan blue,96%-99% BM-MNCs were alive. BM-MNCs were labeled with DAPI blue fluorescence cell linker to detect transplanted cells. DAPI label rate was 100%.BM-MNCs can be cultured in vitro and have spindle shape. The surface antigen CD44 was positive,which is a method for identification of mesenchymal stem cells （MSCs）. Conclusion BM-MNCs are easy to be isolated and cultured in vitro, therefore BM-MNCs have been considered as prospective stem cells in cell transplantation.