Properties of murine embryonic stem cells maintained on human foreskin fibroblasts without LIF
Properties of murine embryonic stem cells maintained on human foreskin fibroblasts without LIF
复制标题
DOI：
10.1002/mrd.20790
复制
发表时间：
2008-04-01
影响因子：
2.5
通讯作者：
Rancourt, D. E.
中科院分区：
文献类型：
--
作者：
Meng, G. L.;Nieden, N. I. Zur;Rancourt, D. E.
在胚胎干（ES）细胞中，白血病抑制因子（LIF）/STAT 3、wnt和nodal/激活素信号传导主要活跃以控制扩增期间的多能性。为了维持多能性，ES细胞通常在不同来源的饲养细胞上培养。小鼠ES细胞通常在小鼠胚胎成纤维细胞（MEF）上培养，其早期衰老并且必须频繁制备。该过程是费力的，并且导致批次变化，这对高通量ES细胞扩增提出了挑战。虽然一些细胞系可以通过外源LIF维持，但这种方法成本高。我们在这里提出了一种新的和廉价的培养方法，用于扩大小鼠ES细胞对人类包皮成纤维细胞（HFF）饲养。在没有LIF的HFF上传代20次后，ES细胞系显示出多能性标记物的正常表达水平，保持正常的核型，并保留了贡献于种系的能力。由于HFF至少62代不会衰老，因此它们提供了大量的饲养者。
In embryonic stem (ES) cells, leukemia inhibitory factor (LIF)/STAT3, wnt and nodal/activin signaling are mainly active to control pluripotency during expansion. To maintain pluripotency, ES cells are typically cultured on feeder cells of varying origins. Murine ES cells are commonly cultured on murine embryonic fibroblasts (MEFs), which senesce early and must be frequently prepared. This process is laborious and leads to batch variation presenting a challenge for high-throughput ES cell expansion. Although some cell lines can be sustained by exogenous LIF, this method is costly. We present here a novel and inexpensive culture method for expanding murine ES cells on human foreskin fibroblast (HFF) feeders. After 20 passages on HFFs without LIF, ES cell lines showed normal expression levels of pluripotency markers, maintained a normal karyotype and retained the ability to contribute to the germline. As HFFs do not senesce for at least 62 passages, they present a vast supply of feeders.