慢性高原病患者骨髓间充质干细胞的体外扩增及鉴定-第三军医大学学报.docVIP

慢性高原病患者骨髓间充质干细胞的体外扩增及鉴定 冀林华，孙 静，王红心，崔森，李占全 青海大学附属医院血液科，青海西宁，810001 【摘要】目的 对慢性高原病（chronic mountain sickness，CMS）患者骨髓间充质干细胞（mesenchymal stem cell，MSCs）进行分离培养扩增及鉴定，以探讨其生物学特性。方法 采用密度梯度离心和贴壁筛选法对15例CMS（诊断符合2004年第六届国际高原医学和低氧生理学术大会颁布慢性高原病青海诊断标准）和15例正常人骨髓MSCs进行分离培养，倒置显微镜下观察MSCs形态、，流式细胞仪行细胞表面抗原检测，检测MSCs生长情况，测定生长曲线。结果 骨髓密度梯度离心法分离骨髓单个核细胞（BMMNC）后培养，骨髓MSC均分离培养成功，贴壁细胞呈梭形；两组骨髓MSCs均表达CD29、CD105和CD13，不表达CD45、CD4、CD8、CD14、CD3、CD34、HLA-DR和CD20。CMS骨髓MSCs增殖能力高于正常骨髓（P0.01），但传代率低于正常骨髓（P0.05），对P3代细胞生长曲线进行观察发现CMS骨髓MSCs生长较正常活跃。结论 CMS患者骨髓MSCs体外可有效分离培养，所培养扩增的细胞成分单一，CMS患者骨髓MSCs具有其特有的生物学特性。 【关键词】慢性，高原病； 细胞培养； 间充质干细胞； 生物学特性 Invitroamplification and Identification of Bone Marrow Mesenchymal Stem Cells from Patients with Chronic Mountain Sickness JI Linhua, SUN Jing, WANG Hongxin, CUI Sen, LI Zhanquan, GE Rili. Department of Hematology, Qinghai University Affiliated Hospital，Qinghai Xining 810001，China Ccorresponding author: JI Linhua, Email: lindaji1@163.com 【Abstract】 Objective To explore the invitro culture approaches,biological function and expression of surface marking of bone marrow mesenchymal stem cells(MSCs) from patients with chronic mountain sickness(CMS). Methods Totally 15 male CMS patients aged 32-45 years and 15 male healthy controls were selected.The diagnosis of CMS was used by the Qinghai CMS score,which established during the VI World Congress in 2004.MSCs were isolated from bone marrow of 15 CMS patients and 15 controls after their agreement by density gradient centrifugation and adhesive culture in vitro. MSCs were identified according to morphology, cell surface antigen profile.The morphological change and the proliferation growth curve of MSCs were observed in primary and passage cultures. The surface makers were detected by flow cytometry(FCM). Results MSCs were successfully isolated from 15 CMS patients and controls.Adherent cells morphology were spindle shape. After flow cytometry, MSCs surface antigen verified that MSCs expressed CD29,CD105,CD13,but did not express CD45