猪脂肪间充质干细胞的原代培养及诱导鉴定

摘要

Objective To establish a method for high yield mesenchymal stem cells collection, as well as a culture method for identifying mesenchymal stem cells from the swine adipose-derived mesenchymal stem cell (ADMSC).Methods Swine ADMSCs were isolated from fat tissue with collagenase , followed by induction of differentiation to osteogenic, adipogenic and chondrogrnic cells. The survival curve of the ADMSC at the 37 ºC and 38 ºC were measured using WST-1Cell Proliferation Assay Reagent.Results ADMSCs isolated with collagenase from swine neck fat tissue generated a stable uniform appearance after the second generation. The passage period was ifve days. ADMSC could differentiate into osteogenic, adipogenic or chondrogrnic cells under different culture conditions. The highest growth rate was achieved at 38 ºC in this study.Conclusion Swine ADMSCs have the potential to differentiate into osteogenic, adipogenic or chondrogrnic cells, and they may be appropriate for transplantation for both research and clinical purpose.%目的：原代培养猪脂肪间充质干细胞（adipose-derived mesenchymal stem cell，ADMSC），探索其适宜培养温度并对其进行诱导鉴定。方法采用胶原酶消化法原代分离培养猪脂肪间充质干细胞，用WST-1法比较猪ADMSC在37℃和38℃条件下的生长曲线，向成骨、成脂和成软骨方向诱导分化ADMSC，鉴定其多能性。结果用胶原酶消化法可以从猪颈部脂肪组织中分离培养出脂肪间充质干细胞，贴壁生长，P2代以后形态均一，稳定增殖，约5天可传代，在一定条件诱导下可以向成骨、成脂或成软骨方向分化，适宜在5%CO2、38℃、饱和湿度条件下培养。结论原代培养的猪脂肪间充质干细胞分离方便、增殖速度快、状态稳定，具有多向分化潜能，是可靠的供体细胞来源。