中国医药
中國醫藥
중국의약
CHINA MEDICINE
2011年
9期
1110-1112
,共3页
宋丹妮%蒋绍艳%史玉朋%常宏
宋丹妮%蔣紹豔%史玉朋%常宏
송단니%장소염%사옥붕%상굉
骨髓间充质干细胞%细胞培养%贴壁培养
骨髓間充質榦細胞%細胞培養%貼壁培養
골수간충질간세포%세포배양%첩벽배양
Bone marrow mesenchymal stem cells%Cell culture%Adherent culture
目的 建立体外分离纯化及培养扩增大鼠骨髓间充质干细胞的方法.方法 应用全骨髓贴壁培养法分离培养大鼠骨髓间充质干细胞,并进行细胞传代培养.倒置显微镜下观察细胞形态及生长特征,测定细胞生长曲线,通过流式细胞仪检测细胞表面标志物,取第3代细胞分别加入成骨、成脂诱导剂并采用碱性磷酸酶染色及Von Kossa染色鉴定成骨能力,以油红O染色鉴定成脂能力.结果 获取的大鼠骨髓间充质干细胞形态呈均一成纤维细胞样,并呈集落样生长.传至第3代细胞纯度可达97%以上,其细胞表型CD29、CD44、CD105、CD166呈阳性表达,CD34、CD80、CD86呈阴性表达.经成骨诱导后细胞碱性磷酸酶染色及Von Kossa染色呈阳性,成脂诱导后细胞油红O染色呈现阳性.结论 应用全骨髓贴壁法可以分离培养出高纯度的大鼠bMSCs,培养的细胞扩增迅速、生物学特性稳定,是一种较为理想的体外分离扩增bMSCs的培养体系.
目的 建立體外分離純化及培養擴增大鼠骨髓間充質榦細胞的方法.方法 應用全骨髓貼壁培養法分離培養大鼠骨髓間充質榦細胞,併進行細胞傳代培養.倒置顯微鏡下觀察細胞形態及生長特徵,測定細胞生長麯線,通過流式細胞儀檢測細胞錶麵標誌物,取第3代細胞分彆加入成骨、成脂誘導劑併採用堿性燐痠酶染色及Von Kossa染色鑒定成骨能力,以油紅O染色鑒定成脂能力.結果 穫取的大鼠骨髓間充質榦細胞形態呈均一成纖維細胞樣,併呈集落樣生長.傳至第3代細胞純度可達97%以上,其細胞錶型CD29、CD44、CD105、CD166呈暘性錶達,CD34、CD80、CD86呈陰性錶達.經成骨誘導後細胞堿性燐痠酶染色及Von Kossa染色呈暘性,成脂誘導後細胞油紅O染色呈現暘性.結論 應用全骨髓貼壁法可以分離培養齣高純度的大鼠bMSCs,培養的細胞擴增迅速、生物學特性穩定,是一種較為理想的體外分離擴增bMSCs的培養體繫.
목적 건입체외분리순화급배양확증대서골수간충질간세포적방법.방법 응용전골수첩벽배양법분리배양대서골수간충질간세포,병진행세포전대배양.도치현미경하관찰세포형태급생장특정,측정세포생장곡선,통과류식세포의검측세포표면표지물,취제3대세포분별가입성골、성지유도제병채용감성린산매염색급Von Kossa염색감정성골능력,이유홍O염색감정성지능력.결과 획취적대서골수간충질간세포형태정균일성섬유세포양,병정집락양생장.전지제3대세포순도가체97%이상,기세포표형CD29、CD44、CD105、CD166정양성표체,CD34、CD80、CD86정음성표체.경성골유도후세포감성린산매염색급Von Kossa염색정양성,성지유도후세포유홍O염색정현양성.결론 응용전골수첩벽법가이분리배양출고순도적대서bMSCs,배양적세포확증신속、생물학특성은정,시일충교위이상적체외분리확증bMSCs적배양체계.
Objective To establish a method of isolating and cultivating rat bone marrow mesenchymal stem cells(BMSCs) in vitro and explore the biological characteristics. Methods BMSCs were isolated and purified for culture in vitro and serial passage by the adherent culture method. The morphological changes of the bMSC were continuously observed under the inverted microscope. The cell growth curve was measured by MTT method. The third generation of bMSCs were detected with flow cytometry(FCM), and incubated in osteogenesis and adipogenesis. Osteogenous potential was assessed by alkaline phosphatase staining and Von Kossa staining. Adipogenic potential was evaluated by Oil red O staining. Results The adherent cells showe spindle shape, polygonal shape and fibroblastcell-like shape and the size of bMSCs were homogeneous. CD29, CD44, CD105 and CD166 memberane antigens could be detected in 97% of the third generation of bMSCs, but CD34, CD80 and CD86 membrane antigens were not detected. Following induction, alkaline phosphatase staining and Oil red O staining produced a strong reaction in cells.Conclusion The whole bone marrow adherent culture is a convenient and efective method to obtain purified bMSCs with strong proliferative ability and capability of multidirectional differentiation.
