小型猪骨髓间充质干细胞的分离和培养

目的建立小型猪骨髓间充质干细胞（mesenchymal stem cells,MSCs）的体外分离和培养方法。方法穿刺小型猪髂后上嵴抽取骨髓,经密度梯度法离心得到骨髓单个核细胞,接种后形成单层贴壁细胞。用形态学方法鉴定培养的MSCs。结果经培养存活的MSCs原代和一代呈纺锤型、多边型或星型。至二代起呈均一纺锤型,似成纤维细胞样,长宽比例约为（2～3）？1。体外培养的原代MSCs8～10d达到融合,传代后仍具有较强的增殖能力。结论小型猪MSCs可在体外长期、稳定培养,其分离、培养体系的建立为基础研究和组织工程技术提供了一个有价值的动物模型。

Isolation and culture of mesenchymal stem cells from miniswine bone marrow

Objective To establish a method of isolation and culture of mesenchymal stem cells（MSCs） from miniswine bone marrow. Method MSCs were isolated from bone marrow by extracting from the posterior superior iliac spine of miniswines and purified by centrifuge and then cultured in single layer in vitro. MSCs quality were evaluated with morphology.Result Survival MSCs appeared morphologically as fusiform and polygon or star in primary and first cultures. All the cultured cells appeared morphologically as fusiform and fibroblast cell-shaped; their length/width ratio was （2~3）：1 from second culture. They were confluent after plating for 8~10 days and showed active proliferation in vitro after passage cultures.Conclusion The MSCs isolated from miniswine bone marrow can be cultured stably for long period in vitro. The establishment of isolating and culturing MSCs of miniswines provided a useful animal model for basic study and tissue engineering.