CD31免疫磁珠分选人脂肪内皮祖细胞的培养及鉴定

发布时间：2018-06-27 04:34

  本文选题：脂肪组织 + 抗原　； 参考：《中国组织工程研究》2017年29期

【摘要】：背景:内皮祖细胞是成熟内皮细胞的前体细胞,可迁移到缺血组织,分化为成熟内皮细胞,发挥血管新生作用。在各种缺血性疾病具有广阔的应用前景,但其生物学特性及鉴定方法仍存在争议。目的:探讨从人脂肪组织中分离培养内皮祖细胞的方法,并进行鉴定,为移植治疗缺血性疾病提供足量的细胞来源。方法:采用酶消化法从人脂肪组织中分离培养出脂肪基质血管细胞(stromal vascular cells,SVF),再通过免疫磁珠分选出CD31~+细胞,体外扩增培养得到内皮祖细胞,并通过细胞形态、细胞生长增殖能力,细胞表型及细胞的功能多方面来完成鉴定。结果与结论:①免疫磁珠分选后的CD31+细胞,体外扩增培养呈典型的铺路石样形态,且其保持一定的生长增殖潜能;②第5代CD31~+细胞流式分析结果显示:CD31、CD34、VEGRR2、CD146和CD133表达量分别为98.84%、97.21%、64.07%、98.42%和2.55%,但几乎不表达造血干细胞表面标志CD45(1.1%);③免疫荧光染色结果表明:CD31~+细胞可摄取乙酰化低密度脂蛋白(Dil-ac-LDL)并可结合荆豆凝集素(FITC-UEA-Ⅰ)。并且,CD31+细胞在Matrigel基质胶中可形成管腔样结构,具有体外成血管能力;④结果提示,通过酶消化法结合免疫磁珠分选法从人脂肪组织中分离培养内皮祖细胞是可行的。
[Abstract]:Background: endothelial progenitor cells are progenitor cells of mature endothelial cells, which can migrate to ischemic tissue, differentiate into mature endothelial cells and play a role in angiogenesis. It has a broad application prospect in various ischemic diseases, but its biological characteristics and identification methods are still controversial. Aim: to investigate the method of isolation and identification of endothelial progenitor cells (EPCs) from human adipose tissue in order to provide sufficient cell sources for transplantation and treatment of ischemic diseases. Methods: (stromal vascular cells were isolated from human adipose tissue by enzyme digestion. CD31 ~ cells were isolated by immunomagnetic beads. Endothelial progenitor cells (EPCs) were obtained by cell morphology and cell proliferation. Cell phenotypes and cell functions are identified in many ways. Results and conclusion the CD31 cells isolated by immunomagnetic beads showed typical paving stone morphology in vitro and maintained a certain growth and proliferation potential. 2The results of flow cytometry of CD31 ~ in the fifth generation showed that the expression levels of CD146 and CD133 of VEGRR2G CD146 and CD133 were 98.84 ~ 97.21% and 64.07% and 2.55%, respectively, but hardly expressed CD45 (1.1%) of CD45 (1.1%) on the surface of hematopoietic stem cells. The agglutinin (FITC-UEA- 鈪，

本文编号：2072688