人羊膜上皮细胞干细胞特性及向胰岛素分泌细胞分化的研究.pdfVIP

·1484 · 南方医科大学学报（J South Med Univ ） 2011;31(9) ·基础研究· 人羊膜上皮细胞干细胞特性及向胰岛素分泌细胞分化的研究 王 建，彭 琳，卢光琇 中南大学生殖与干细胞工程研究所/人类干细胞国家工程研究中心，湖南 长沙 410078 摘要：目的建立人羊膜上皮细胞（hAECs ）的分离及培养方法，探讨其向胰岛素分泌细胞分化的能力。方法 观察不同培 养方法对hAECs 生长倍增时间的影响以优化其培养体系；通过细胞免疫荧光法检测不同培养代数细胞多能干细胞的表 面标记；通过体外添加诱导因子的方法，探讨诱导hAECs 向胰岛素分泌细胞分化的潜能。结果 在10 ng 表皮生长因子培 养下，hAECs 可长期传代。hAECs 表达胚胎干细胞表面标记如阶段特异性胚胎抗原4 等；采用悬浮培养法诱导分化，可检 测到胰岛发育、胰岛功能基因如胰腺十二指肠同源盒、神经源素3、同源异形盒转录因子6、胰岛素、胰高糖素的表达，免疫 组化检测细胞可以表达胰岛素，并且随着外界葡萄糖浓度升高，胰岛素分泌显著增加。结论 建立了人羊膜上皮细胞的 分离和培养方法（不添加动物血清）；在体外可诱导分化为胰岛素分泌细胞。 关键词：人羊膜上皮细胞；胰岛素分泌细胞；横向分化 中图分类号：R335.6; R34 文献标志码：A 文章编号：1673-4254（2011）09-1484-04 Stem cell characteristics and islet differentiation potential of human amniotic epithelial cells WANG Jian, PENG Lin, LU Guang-xiu Institute of Reproductive and Stem Cell Engineering, Central South University; National Center of Human Stem Cell Research and Engineering, Changsha 410078, China Abstract: Objective To establish a method for isolating and culturing human amniotic epithelial cells (hAECs) in a serum-free medium and investigate their transdifferentiation ability into islet-like cells. Methods The culture condition of hAECs was optimized using DMEM with different supplements. The genetic stability of the tenth-passage cells was assessed by chromosome analysis and G-banding method. The stem cell characteristics of the cells were identified by examination of the surface markers using immunofluorescence methods. The endocrine-related genes and hormones of the cells were tested after induced differentiation into islet-like cells. Results The hAECs allow stable passaging in the prese