无饲养细胞无外源因子条件下高克隆密度维持人类胚胎干细胞自身未分化状态(英文)

Maintenance of undifferentiated state of human embryonic stem cells in chemical defined medium at high clone density without exogenous cell factors

目的:建立人类胚胎干细胞(human embryonic stem cells,hESCs)无饲养细胞无外源性细胞因子体外培养体系。方法:比较hESCs克隆种植密度对其无饲养细胞无外源性细胞因子体外培养的影响,以及克隆种植密度对hESCs培养体系骨形蛋白(bone morphology protein,BMP)样诱导活性的影响。结果:在没有外源性细胞因子条件下,高克隆密度(34克隆/cm2)可以维持hESCs处于未分化状态,而低克隆密度不能维持hESCs处于未分化状态。同时,NB体系BMP样诱导活性可以被hESCs克隆种植密度所改变,高hESCs克隆密度时,培养基具有高的BMP样诱导活性。结论:高hESCs克隆种植密度可以改变自身生长的微环境,可以维持自身未分化状态,这为阐明hESCs自身未分化状态机制和简化hESCs无饲养培养体系提供了新的思路。

Objective To establish human embryonic stem cells （ hESCs） feeder-independent and cell factor-free culture system. Methods Effect of high and low clone densities of hESCs culture was compared and impact of the clone densities to hESCs culture media was analyzed. Results HESCs could maintain their undifferentiated states at high clone density （ 34 clones/cm2） without cell factors. At the same time,the bone morphology protein （ BMP）-like induction of N2 and B27 supplements （ NB） medium could be modulated by the clone density,and high level of BMP-like induction was accompanied by high clone density. Conclusion High clone density of hESCs can change the environments by them-selves to maintain the undifferentiated states,which provides a new clue to explore the mechanism of undifferentiated states of hESCs and simplify the culture medium.