摘要
背景:内皮祖细胞可从外周血与脐血中获取,是修复各种疾病所致损伤的血管内皮细胞不可或缺的细胞来源。目的:比较人外周血与脐血来源的内皮祖细胞经体外培养后生物学特性的差异。方法:通过密度梯度离心法和6%羟乙基淀粉结合密度梯度离心法分别分离人外周血与脐血中的单个核细胞,分别设为脐血源组和外周血源组,计数各组单个核细胞数量,按1.0×106/cm2接种于大鼠尾胶包被的培养皿中,用内皮细胞培养基进行诱导,共培养7 d。结果与结论:外周血与脐血体外培养分离出内皮祖细胞具有类似的形态学特征。光学显微镜下观察,随着培养天数的增加,大多数细胞由早期的贴壁圆形转变为梭形。外周血源组内皮祖细胞有细胞集落形成,脐血源组可见梭形细胞自行排列生长为典型的线样结构。锥虫蓝染色及绘制细胞生长曲线后发现,外周血源组单个核细胞及内皮祖细胞数量、内皮祖细胞活率及增殖能力均低于脐血源组(P<0.05)。外周血源组和脐血源组内皮祖细胞在接种后第3天增殖速度达到峰值,在随后的培养中细胞增殖呈衰减态。流式细胞仪及免疫荧光染色检测结果显示,外周血源组和脐血源组内皮祖细胞均可表达具有内皮祖细胞表型的CD133、CD34和血管内皮细胞因子受体2表面标志物,两组既摄取Dil标记乙酰化低密度脂蛋白,也能标记体外内皮祖细胞的标志物荆豆凝集素Ⅰ。结果证实,脐血来源的内皮祖细胞与外周血来源的内皮祖细胞生物学特性相近,脐血来源的内皮祖细胞增殖能力更强。
BACKGROUND： Endothelial progenitor coils from the peripheral blood and umbilical cord blood are an essentia source to repair vascular endothelial cells damaged by vadous diseases. OBJECTIVE： To compare the biological characteristics of endothelial progenitor cells from peripheral blood and umbilical cord blood in vitro. METHODS： Mononuclear cells were isolated from the umbilical cord blood and peripheral blood with density gradient contrifugation method and 6% hydroxyethyl starch precipitation combined with density gradient contrifugation method, respectively. Mononuclear coUs were counted. Then the coUs were implanted on the culture plate pre-paved with rat tail collagen at the concentration of I x 106/cm2, and cultured in an endothelial col culture medium for 7 days. RESULTS AND CONCLUSION： Isolated and cultured endothelial progenitor cells from the peripheral blood and umbilical cord blood had similar morphological characteristics in vitro. Under the optical microscope, with theincrease of culturing days, most adherent calls were changed from round to spindle-shaped. Peripheral blood endothelial progenitor cells had cell colony formation, and spindle calls from the umbilical cord blood arranged typically in a line structure. After trypan blue staining and drawing of cell growth curves, the number of mononuclear cells and endothelial progenitor cells, survival rate and proliferative ability of endothelial progenitor cells from the peripheral blood were all lower than those from the umbilical cord blood （P 〈 0.05）. At day 3 after incubation, the proliferation of endothelial progenitor calls from the peripheral blood and umbilical cord blood reached the peak, and then showed a decreased tendency. Flow cytometry and Immunofluorescance staining showed that endothelial progenitor cells from the peripheral blood and umbilical cord blood could express CD34, CD133, and vascular endothelial cell factor receptor 2. The endothelial progenitor cells from the peripheral blood and umbilical cord blood could swallow acatylated low density lipoprotein and be combined with ulex europaeus agglutinin I. The results confirmed that the endothelial progenitor cells from the peripheral blood and cord blood have similar biologicla characteristics, but the proliferation ability of endothelial progenitor cells from the cord blood is higher.
出处
《中国组织工程研究》
CAS
CSCD
2013年第45期7911-7917,共7页
Chinese Journal of Tissue Engineering Research
基金
新疆维吾尔自治区自然科学基金(2011211A075)~~
