用于人类脂肪干细胞的纯化和体外长期培养的聚合物涂层

发布时间：2018-06-24 20:43

本文选题：聚合物涂层 + 人类脂肪干细胞　；参考：《高等学校化学学报》2017年10期

【摘要】：采用紫外固化法制备了基于丙烯酸酯类水凝胶的聚合物涂层(PC),并用X射线光电子能谱(XPS)、水接触角(WCA)和原子力显微镜(AFM)分别对PC进行了化学组成和表面性能的表征.在PC表面进行了人类脂肪干细胞(h ASC)的体外长期培养扩增,得到的第3代细胞的生物学表征结果表明,干细胞在PC表面能正常黏附生长,流式细胞仪检测发现干细胞对特征标记物CD49d,CD73,CD105的阳性显性比例较高,对HLA-DR和CD31几乎不显性,说明扩增的干细胞具有h ASC特征.对PC上扩增的干细胞进行诱导分化,并用油红O、茜素红和阿利新蓝分别进行染色分析,结果表明,该干细胞保留了h ASC的多能特性:能分化为成脂、成骨和成软骨细胞.含有单体甲基丙烯酰氧乙基三甲基氯化铵(DMC)、甲基丙烯酸环己酯(CHMA)和甲基丙烯酸-2-(二乙氨基)乙酯(DEAEMA)的PC2(质量比为3∶1∶2)在用于h ASC体外长期培养时,比其它PC和TCP更有利于细胞的黏附和增殖,纯化细胞,保持其多能性.实时荧光定量PCR(RT-q PCR)的分析表明PC2上得到的细胞更容易向成骨和成软骨细胞分化.
[Abstract]:Polymer coatings (PC) based on acrylate hydrogels were prepared by UV curing method. The chemical composition and surface properties of PC were characterized by X-ray photoelectron spectroscopy (XPS), water contact angle (WCA) and atomic force microscope (AFM). Human adipose stem cells (hASC) were cultured and amplified on PC surface for a long time in vitro. The results of biological characterization of the third passage cells showed that the stem cells could grow normally on the PC surface. The results of flow cytometry showed that the stem cells had a high percentage of positive dominance to CD49dU CD73 and CD105, but almost no dominance to HLA-DR and CD31, indicating that the expanded stem cells had hASC characteristics. The stem cells amplified on PC were induced to differentiate and stained with oil red O, alizarin red and alisin blue respectively. The results showed that the stem cells retained the pluripotent characteristics of hASC: they could differentiate into fat-forming, osteogenic and chondrogenic cells. PC2 containing monomers methacryloxy ethyl trimethylammonium chloride (DMC), cyclohexyl methacrylate (CHMA) and diethylaminoethyl methacrylate (DEAEMA) (with a mass ratio of 3:1:2) was used in long-term culture of hASC in vitro. Compared with other PC and TCP, it is more favorable to cell adhesion and proliferation, purify cells and maintain their pluripotency. Real-time fluorescent quantitative PCR (RT-q PCR) analysis showed that PC2 cells were more likely to differentiate into osteoblasts and chondroblasts.
【作者单位】：常州大学材料科学与工程学院;常州大学生物医学工程与健康科学研究院;江苏和泽生物科技有限公司;常州市第二人民医院;School
【基金】：国家自然科学基金(批准号:21374012)资助~~
【分类号】：Q813.1;R318.08

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