Wang Jian, Peng Lin, Lu Guang-xiu
Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha , China.
Nan Fang Yi Ke Da Xue Xue Bao. 2011 Sep;31(9):1484-7.
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.
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.
The hAECs allow stable passaging in the presence of 10 ng epidermal growth factor (EGF) in the culture medium. After 10 passages, the cells maintained a normal karyotype and G-banding profile. The hAECs expressed many multi-potent stem cell markers, including SSEA4, TRA-1-60, and TRA-1-81. After induced differentiation, the endocrine-related genes were expressed in the islet-like cells, including PDX1, ngn3, insulin and glucagon. Insulin secretion increased in the differentiated islet-like cells in response to high glucose exposure.
We established a method for isolating and expanding the hAECs in a serum-free medium. hAECs possess stem cell characteristics and can be induced to differentiate into islet-like cells in vitro.
建立在无血清培养基中分离培养人羊膜上皮细胞（hAECs）的方法，并研究其向胰岛样细胞的转分化能力。
使用添加不同成分的DMEM优化hAECs的培养条件。通过染色体分析和G显带法评估第十代细胞的遗传稳定性。采用免疫荧光法检测细胞表面标志物以鉴定细胞的干细胞特性。将细胞诱导分化为胰岛样细胞后，检测其内分泌相关基因和激素。
在培养基中存在10 ng表皮生长因子（EGF）的情况下，hAECs能够稳定传代。传代10次后，细胞保持正常的核型和G显带模式。hAECs表达多种多能干细胞标志物，包括SSEA4、TRA-1-60和TRA-1-81。诱导分化后，胰岛样细胞中表达内分泌相关基因，包括PDX1、ngn3、胰岛素和胰高血糖素。分化后的胰岛样细胞在高糖刺激下胰岛素分泌增加。
我们建立了在无血清培养基中分离和扩增hAECs的方法。hAECs具有干细胞特性，并且在体外可被诱导分化为胰岛样细胞。
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