Effect of Human Feeders on Growth of Human Embryonic Stem Cells / 中山大学学报(医学科学版)

ABSTRACT

[Objective] The aim was to choose the best feeder layer by observing the effects of various human feeders supporting human embryonic stem cells (hESCs), and to probe the correlation between the levels of basic fibroblast growth factor (bFGF) secreted by feeders and the growth of the hESCs. [Methods] The primary cells from various tissues were cultured, including foreskin, stromal endometrium, villus, adult fallopian tubal, fetal skin, fetal muscle and mouse embryonic fibroblasts (MEFs). The hESCs were transferred to various feeders, and then the best condition was probed, which was based on the feeder density and the time of mitomycin-C acting on the feeder. Comparing the characteristics of the hESCs, the best feeder was chosen of all kinds of feeders from various tissues that support the hESCs. The level of bFGF secreted by various feeders was detected using ELISA. [Results] All of tested feeders could support the hESCs growth for over 10 passages in the culture, especially the foreskin and the adult fallopian tubal. The density of feeders was related with the morphology and the differentiation rate of the hESCs. According to the characteristics of feeder, the feeder ranking was as follows foreskin, stromal endometrium, villus, adult fallopian tubal, fetal skin and fetal muscle. Based on the characteristics of the hESCs, the order of feeders was foreskin, adult fallopian tubal, stromal endometrium, villus, fetal muscle and fetal skin. The levels of bFGF (pg·10~(-5)·mL~(-1)) secreted by various feeders were as follows adult fallopian tubal (13.23±3.39), foreskin (1.99±0.17), villus (1.40±0.17), fetal muscle (2.02 ±1.59), stromal endometrium (0.38±0.28), and fetal skin (0.29±0.29). [Conclusion] The foreskin and the adult fallopian tubal could support the hESCs better than others though all of them could;do, especially the, foreskin. The bFGF that secreted by the adult fallopian tubal was the highest of all. The correlation was not obvious .between the levels of bFGF secreted by feeders and the growth of the hESCs.