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从大西洋鳕鱼(Gadus morhua)中分离的胚胎干细胞和阶段特异性转录因子 ac-Pou2 的发育表达。

Embryonic stem cells isolated from Atlantic cod (Gadus morhua) and the developmental expression of a stage-specific transcription factor ac-Pou2.

机构信息

National Institute of Nutrition and Seafood Research (NIFES), Post Box 2029 Nordnes, 5817, Bergen, Norway.

出版信息

Fish Physiol Biochem. 2010 Dec;36(4):1029-39. doi: 10.1007/s10695-010-9381-z. Epub 2010 Feb 4.

Abstract

The establishment of embryonic stem cell cultures and the identification of molecular markers for undifferentiated embryonic stem cells (ESC) as well as differentiated cells types will open new opportunities in the study of developmental biology and for developing embryonic in vitro models of the ecologically and economically important fish specie Atlantic cod (Gadus morhua). We report here that cod blastula cells express a Class V POU gene known to be highly expressed in embryonic cell populations of vertebrates. The cod transcript, designated Atlantic cod-Pou2 (ac-Pou2), can be used as a genetic marker for cod blastula cells in vivo and in vitro. Using a quantitative real-time PCR approach, we found that the ac-Pou2 transcript was downregulated before the egg reached the stage of gastrulation, the starting point of extensive cell differentiation. We also demonstrate the culturing of ESC isolated from cod blastula stage eggs. The cod ESC exhibited in vitro characteristics of pluripotency described for both mammalian ESC and fish ES-like cells (medaka, zebrafish, seabream, sea perch and rainbow trout). Cod ESC in culture expressed ac-pou2, differentiated spontaneously and had the ability to form embryoid bodies following retinoic acid treatment. The ESC could also be directed to differentiate.

摘要

胚胎干细胞培养的建立和未分化胚胎干细胞(ESC)以及分化细胞类型的分子标记的鉴定将为发育生物学的研究和生态经济重要鱼类大西洋鳕鱼(Gadus morhua)的体外胚胎模型的开发开辟新的机会。我们在这里报告,鳕鱼囊胚细胞表达一种已知在脊椎动物胚胎细胞群体中高度表达的 Class V POU 基因。鳕鱼转录本,指定为 Atlantic cod-Pou2 (ac-Pou2),可作为体内和体外鳕鱼囊胚细胞的遗传标记。使用定量实时 PCR 方法,我们发现 ac-Pou2 转录本在卵达到广泛细胞分化起始点原肠胚形成阶段之前下调。我们还证明了从鳕鱼囊胚期卵中分离的 ESC 的培养。鳕鱼 ESC 表现出与哺乳动物 ESC 和鱼类 ES 样细胞(日本青鳉、斑马鱼、真鲷、黑鲷和虹鳟)描述的体外多能性特征。在培养中,ESC 表达 ac-pou2,自发分化,并在视黄酸处理后具有形成胚状体的能力。ESC 也可以被定向分化。

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