摘要
目的:探讨骨髓间充质干细胞体外扩增及其向内皮样细胞定向分化条件。方法:无菌条件下采集兔骨髓,肝素抗凝,经淋巴细胞分层液(相对密度为1.077)密度梯度离心分离骨髓单个核细胞,通过贴壁培养法获得骨髓间充质干细胞(bonemarrow-mesenchymalstemcell,MSC),然后在内皮细胞生长环境中进行诱导分化,观察细胞的体外生长特点及其生化特性。结果:MSC每扩增一代,细胞数量增加5～8倍,7.65×103个原代MSCs体外扩增3代即获得1.26×108个细胞。特定条件下贴壁细胞延展呈梭形,形成细胞簇、索状结构及“铺路石”样结构,扩增细胞免疫组化证实表达CD31和VIII因子相关抗原(factorVIIIrelatedantigenvonWille-brandfactor,vWF),具有吞噬乙酰化低密度脂蛋白的功能。结论:MSC扩增能力强,在体外能诱导其向内皮细胞方向分化。
AIM:To explore the conditions of isolation,expansion and differentiation of bone marrow mesenchymal stem cells(MSCs) into endothelium like cells in vitro. METHODS:Rabbit bone marrow was sterilely collected from the iliac crest with heparin against coagulation.Bone marrow mononuclear cells were isolated by gradient density centrifugation with lymphocyte separation medium(relative density was 1.077).MSCs were then obtained and cultured in endothelial cell complete medium.Growth property and biochemistry characteristics of expanded cells were detected. RESULTS:The number of MSCs increased by 5-8 times with each expanded passage,and 7.65×103 primary MSCs were expanded to reach a number of 1.26×108 in three passages.Attached cells,spindle cells,cord like structure and cell clusters,cobblestone like structure were observed within the period of culture.The results of immunochemistry showed that expanded cell exhibited typical CD31 and factor VIII related antigen von Willebrand factor(vWF),and had the function of phagocytosis acetylated low density lipoprotein(ac LDL). CONCLUSION:Bone marrow MSCs have the capability of differentiating into endothelial cells in vitro,which can be induced by strengthened amplification.
出处
《中国临床康复》
CSCD
2004年第18期3508-3509,F003,共3页
Chinese Journal of Clinical Rehabilitation
