人脐血清对人脐带血间充质干细胞生物学特征与衰老的影响  

Effects of human umbilical cord blood serum on the biological characteristics and senescence of human umbilical cord blood mesenchymal stem cells

作　　者：王爽[1] 范振海[1] 刘祖林[1] 王钰莹[1] 方宁[1] 喻皇飞[1] 刘金伟[1] 余丽梅[1] 

机构地区：[1]遵义医学院附属医院贵州省细胞工程重点实验室,遵义563003

出　　处：《重庆医科大学学报》2014年第5期582-586,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:30860388);遵义市重大科技项目培养与科技人才培养计划资助项目[编号:遵科培(2009)02号]

摘　　要：目的:探讨人脐血清(human umbilical cord blood serum,CBS)对体外培养人脐血间充质干细胞(human umbilical cord blood mesenchymal stem cells,hUCB-MSCs)基本生物学特征的影响。方法:密度梯度离心分离脐带血单个核细胞,以含3%CBS加7%胎牛血清(fetal bovine serum,FBS)(CBS组)或10%FBS(FBS组)的完全培养基培养hUCB-MSCs,流式细胞术检测细胞表面分子与细胞周期,显微镜下测定细胞表面积和表达β-半乳糖苷酶细胞百分率,MTT法检测光密度值、绘制生长曲线。结果:CBS组和FBS组均表达CD44、CD105、CD90、CD73,不表达CD45、CD34、CD11b、CD19和人白细胞DR抗原(human leukocyte antigen DR,HLA-DR),但与FBS组比较,CBS组表达CD90细胞百分率更低,G0/G1期细胞比例较少,细胞增殖指数较高(P<0.05)且细胞表面积也明显更小(P<0.05)。CBS组生长曲线较FBS组左移,峰值更高,细胞倍增时间48.37 h,FBS组则为54.43 h。FBS组β-gal染色阳性细胞百分率明显高于CBS组(P<0.05)。结论:部分CBS合用FBS培养hUCB-MSCs能更好地促进细胞增殖,并更好地保持hUCB-MSCs的基本生物学特征和延缓细胞的衰老。Objective：To explore effects of human umbilical cord blood serum on the basic biological characteristics and senescence of human umbilical cord blood mesenchymal stem cells（hUCB-MSCs）. Methods：Mononuclear cells were isolated by density gradient centrifugation from human umbilical cord blood. Two groups of hUCB-MSCs were cultured using different complete mediums,one consisting of 3% cord blood and 7% fetal bovine serum（CBS group）,the other one consisting of 10% fetal bovine serum（FBS group）. Cell surface moleculars and cell cycle were determined by flow cytometry. Then cell surface area and percentage of positive cells expressing β-galactosidase were detected. Absorbance value was measured by MTT methods,and cell growth curve was drawn. Results： hUCB-MSCs expressed CD44,CD105,CD90,CD73 and vimentin in both groups,but not CD45,CD34,CD11b,CD19 and human leukocyte antigen DR（HLA-DR）. Percentage of CD90 expression in the 7th passage hUCB-MSCs was significantly lower in CBS group than in FBS group. Percentage of cells in G0/G1 phase was significantly lower and proliferation index was significantly higher. Cell surface area was significantly smaller in CBS group than in FBS group（P＆lt;0.05）. Growth curve of CBS group was shifted to the left with a higher peak. Cell doubling time was 48.37 h in CBS group,but 54.43 h in FBS group. Percentage of β-galactosidase positive cells was higher in FBS than in CBS group. Conclusions：hUCB-MSCs proliferation is promoted by combining partial human umbilical cord blood serum with fetal bovine serum with better maintenance of basic biological characteristic and delayed cell aging.

关 键 词：人脐血清 脐带血间充质干细胞 生物学特征 衰老 

分 类 号：Q813.1[生物学—生物工程]