摘要
目的探索人骨髓间质干细胞(hMSC)的体外分离、扩增、冻存、复苏的方法。方法无菌条件下抽取骨外伤髓内钉固定手术髓腔内容物,应用Ficoll-Paque分离液分离hMSC,通过传代扩增细胞,“慢冻速融”进行细胞冻存、复苏;流式细胞仪检测hMSC的表面标志。结果hMSC容易分离获取、体外扩增迅速、冻存复苏简便,复苏后细胞形态保持不变。流式细胞仪检测结果显示,CD34、CD45阴性,CD29及CD71、CD90阳性表达率分别为96.9%,99.1%及4.1%。结论hMSC库有望建立,为组织工程、细胞移植的研究提供种子细胞。
Objective To establish a method for isolation,cultivation, freeze and resuscitation of human mesenchymal stem cells（hMSC） from human bone marrow. Methods hMSC were separated from human bone marrow with Ficoll-Paque reagent, expanded in vitro, then were freezing depot and resuscitation through rapid melting. To detect the surface antigens, the labeled cells were analyzed with FACScan flow cytometer. Results hMSC were easily isolated from human bone marrow, expanded quickly in vitro, and were easy freeze and resuscitation. The surface antigens in these expanded hMSC were uniformly positive for CD 29, CD 71 CD 90, while the CD 34, CD 45were negative. Conclusion The bank of hMSC is incipiently established and can provide eligible seed cell for constitutive engineering and cell therapy.
出处
《福建医科大学学报》
2007年第2期176-178,共3页
Journal of Fujian Medical University
基金
教育部科学技术研究重点项目(01058)
福建省科技厅科研基金资助项目(2002Y014)
关键词
骨髓细胞
质干细胞
低温保存
组织工程
细胞
培养的
bonemarrow cells
mesenchymal stem cells
cryopreservation
tissue engineering
cells, cullured
