骨质颗粒分离培养间充质细胞的实验研究

An experimental study of isolation and culture of mesenchymal stem cells

目的:为探索间充质干细胞更广泛的来源。方法:选择了正常人骨髓,急性粒细胞性白血病缓解期患者骨髓、再生障碍性贫血患者骨髓、脐带血和骨质颗粒等5种标本来源。采用贴壁法和特殊的干细胞培养基/间充质干细胞刺激因子分离、纯化MSCs,观察比较不同来源MSCs的体外生长特性,流式细胞术检测培养细胞的CD105。结果:正常人骨髓、急性粒细胞性白血病缓解期患者骨髓和骨质颗粒标本均能培养出MSCs,从骨质颗粒得到的MSCs数量较多。脐带血和再生障碍性贫血患者骨髓标本没有培养出MSCs。培养的MSCs均一地表达CD105。结论:在体外培养人类正常骨髓和急性粒细胞性白血病缓解期的骨髓均可以得到MSCs,两种样本分离培养得到的MSCs在形态学、表面标志、增殖特性等方面没有显著差异。从人类骨质颗粒得到的MSCs的生长从数量上优于从骨髓中获得的MSCs。

Objective ：different sources of MSCs isolated from ,humans＇ body . Methods ： Isolation, culture and induced differentiation of MSCs： Samples of bone marrow of normal humans, acute myeloblastic leukemia （AML） and aplastic anemia patients, and cord blood and bone microparticles were used to isolate MSCs. By means of adherence method and specific media MSCs were purified. The growth nature of MSCs from various sources was investigated and surface markers of MSCs were detected by flow cytometry （FCM）. Results： MSCs were expanded from human normal and AML bone marrow and bone microparticles. The highest quantity of MSCs was harvested from bone microparticles. No MSCs could be obtained from cord blood and bone marrow of aplastic anemia patients. MSCs positively expressed CD29 and CD105, but not CD34 and CD45, detected by FCM. Conclusions：MSCs can be obtained from bone marrow of normal humans and AML patients. MSCs from both sources are similar on morphology, cell surface markers and growth nature. MSCs from human bone microparticles are better than those from bone marrow in term of quantity.