生物
重编程
同源盒蛋白纳米
SOX2
诱导多能干细胞
体细胞核移植
体细胞
胚胎干细胞
细胞生物学
胚状体
科斯尔
干细胞
转基因
遗传学
胚泡
胚胎
基因
胚胎发生
作者
Sergio D. German,Keith Campbell,Elisabeth M. Thornton,Gerry McLachlan,Dylan Sweetman,Ramiro Alberio
出处
期刊:Cellular Reprogramming
[Mary Ann Liebert]
日期:2015-02-01
卷期号:17 (1): 19-27
被引量:21
标识
DOI:10.1089/cell.2014.0071
摘要
Induced pluripotent stem cells (iPSCs) share similar characteristics of indefinite in vitro growth with embryonic stem cells (ESCs) and may therefore serve as a useful tool for the targeted genetic modification of farm animals via nuclear transfer (NT). Derivation of stable ESC lines from farm animals has not been possible, therefore, it is important to determine whether iPSCs can be used as substitutes for ESCs in generating genetically modified cloned farm animals. We generated ovine iPSCs by conventional retroviral transduction using the four Yamanaka factors. These cells were basic fibroblast growth factor (bFGF)- and activin A-dependent, showed persistent expression of the transgenes, acquired chromosomal abnormalities, and failed to activate endogenous NANOG. Nonetheless, iPSCs could differentiate into the three somatic germ layers in vitro. Because cloning of farm animals is best achieved with diploid cells (G1/G0), we synchronized the iPSCs in G1 prior to NT. Despite the cell cycle synchronization, preimplantation development of iPSC-NT embryos was lower than with somatic cells (2% vs. 10% blastocysts, p<0.01). Furthermore, analysis of the blastocysts produced demonstrated persistent expression of the transgenes, aberrant expression of endogenous SOX2, and a failure to activate NANOG consistently. In contrast, gene expression in blastocysts produced with the parental fetal fibroblasts was similar to those generated by in vitro fertilization. Taken together, our data suggest that the persistent expression of the exogenous factors and the acquisition of chromosomal abnormalities are incompatible with normal development of NT embryos produced with iPSCs.
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