摘要
目的探讨将人脂肪组织来源CD34+/CD31-细胞诱导为内皮细胞的可行性。方法脂肪组织来自5例接受脂肪抽吸术的患者,酶消化法获取原代细胞,细胞扩增培养至第二代,应用免疫磁珠仪分选CD34+/CD31-细胞。将此群细胞用VEGF及bFGF进行诱导。流式细胞仪检测分选前后细胞纯度;免疫细胞荧光检测细胞vWF和CD31的表达;荧光显微镜观察细胞摄取低密度脂蛋白的功能;三维培养观察细胞形成血管样结构的能力。结果分选前后CD34+/CD31-细胞比例分别为(12.57±1.83)%和(96.05±6.73)%;细胞诱导10d后呈现内皮细胞典型的"铺路石"样形态,vWF、CD31表达阳性,细胞具有摄取低密度脂蛋白的功能,三维培养条件下形成"树枝样"分叉结构。结论人脂肪组织来源的CD34+/CD31-细胞能够诱导分化为成熟内皮细胞。
Objective To investigate the feasibility of inducing CD34＋/CD31- cells isolated from human adipose tissue into endothelial cells. Methods Subcutaneous adipose tissue was obtained from 5 individuals undergoing plastic surgery by lipoaspiration. The CD34 ＋/CD3＋ population of P2 cells were sorted by MACS, and then were induced by VEGF and bFGF. FACS analysis was performed to detect purity of cells before and after isolation. Immunofluorescence was usod to observe endothelial markers vWF and CD34 - Fluorescence micrascope was smployed to obsen,e the fanctiun of taking up low - density lipoprotein by endothelial cells. Throe - dimensional culture was used to determine the ability of forming capillary - like struetum. Results FACS aualysis showed that the ratio of CD34 ＋/CD31 - calls were （ 12.57 ± 1.83 ） % and （96.05 ± 6.73 ） % respectively before and after isolation （p 〈 0.05）. The induced cells exhibited cobblestone morphology, similar to that of the endothelial cells. The induced cells expressed vWF, CD31 and took up LDL. The formation of ＂branch - like＂ structure confirmed their functional aetivity. Conclusion The CD34＋/CD31- cells derived from human adipose can be induced into mature endothelial cells.
出处
《浙江临床医学》
2008年第1期13-14,共2页
Zhejiang Clinical Medical Journal
关键词
人脂胁组织
内皮细胞
诱导
分化
Human adipose tissue Endothelial cell Induce Diffierentiation
