發(fā)布時間：2018-06-27 04:34

  本文選題：脂肪組織 + 抗原　； 參考：《中國組織工程研究》2017年29期

【摘要】：背景:內(nèi)皮祖細(xì)胞是成熟內(nèi)皮細(xì)胞的前體細(xì)胞,可遷移到缺血組織,分化為成熟內(nèi)皮細(xì)胞,發(fā)揮血管新生作用。在各種缺血性疾病具有廣闊的應(yīng)用前景,但其生物學(xué)特性及鑒定方法仍存在爭議。目的:探討從人脂肪組織中分離培養(yǎng)內(nèi)皮祖細(xì)胞的方法,并進行鑒定,為移植治療缺血性疾病提供足量的細(xì)胞來源。方法:采用酶消化法從人脂肪組織中分離培養(yǎng)出脂肪基質(zhì)血管細(xì)胞(stromal vascular cells,SVF),再通過免疫磁珠分選出CD31~+細(xì)胞,體外擴增培養(yǎng)得到內(nèi)皮祖細(xì)胞,并通過細(xì)胞形態(tài)、細(xì)胞生長增殖能力,細(xì)胞表型及細(xì)胞的功能多方面來完成鑒定。結(jié)果與結(jié)論:①免疫磁珠分選后的CD31+細(xì)胞,體外擴增培養(yǎng)呈典型的鋪路石樣形態(tài),且其保持一定的生長增殖潛能;②第5代CD31~+細(xì)胞流式分析結(jié)果顯示:CD31、CD34、VEGRR2、CD146和CD133表達(dá)量分別為98.84%、97.21%、64.07%、98.42%和2.55%,但幾乎不表達(dá)造血干細(xì)胞表面標(biāo)志CD45(1.1%);③免疫熒光染色結(jié)果表明:CD31~+細(xì)胞可攝取乙�；兔芏戎鞍�(Dil-ac-LDL)并可結(jié)合荊豆凝集素(FITC-UEA-Ⅰ)。并且,CD31+細(xì)胞在Matrigel基質(zhì)膠中可形成管腔樣結(jié)構(gòu),具有體外成血管能力;④結(jié)果提示,通過酶消化法結(jié)合免疫磁珠分選法從人脂肪組織中分離培養(yǎng)內(nèi)皮祖細(xì)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，

本文編號：2072688