摘要
目的 本实验旨在建立一套体外分离培养人骨髓基质干细胞 (humanbonemarrowstemcellHMSCs)的方法 ,观察HMSCs的形态和掌握其生长规律 ,找到一种最佳的培养环境。方法 穿刺抽取成人骨髓约 2 0ml,通过离心法获取HMSCs,体外培养并传代 ,观察原代及传代细胞的细胞形态及生长规律 ,第 4代细胞用矿化液连续培养 30天 ,在倒置显微镜下观察矿化结节形成情况。结果 体外利用骨髓基质细胞培养液成功培养了HMSCs ,并连续传代 5～ 7代。利用第 4代细胞用矿化液连续培养 30天 ,矿化结节形成 ,在细胞间有钙盐沉积。结论 HMSCs可以在体外培养成功 ,并可定向诱导为成骨细胞 。
Objective To create the model of human bone marrow stem cell (HMSCs) separation and culture in vitro.Methods Adult bone marrow about 20ml were drawed by puncture.After centrifuging,HMSCs were harvested,then were culutivated and transfered.The growth and the characteristics of HMSCs were observed through cell morphological survey,cell growth and proliferation and ALP activity.The cells of fourth passage were cultured continiously in mineral condition for 30 days,the mineral nodules were oberseved by Von-Kossa staining.Results HMSCs were cultured in vitro and could be transfered for 5～7 passages steadly.The cells of fourth passage were ALP positive.When culured up to 30 days Von-Kossa staining showed there was calcium deposition in the nodules.Conclusion The HMSCs were cultured succesfully in vitro and can be induced to osteoblast cells,which will hopelully become excellent seeding-cells for bone tissue engineering.
出处
《现代口腔医学杂志》
CAS
CSCD
2002年第5期385-387,T001,共4页
Journal of Modern Stomatology
基金
国家自然科学基金 (编号 :3 9779762 )
国家"973"子课题口腔颌面骨组织工程 (编号 :G19990 5 43 0 8)
第四军医大学创新工程 (编号 :CX99F0 13 )资助
