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首页> 外文期刊>BMC Biotechnology >Retinoic acid promotes expression of germline-specific genes in chicken blastoderm cells by stimulating Smad1/5 phosphorylation in a feeder-free culture system
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Retinoic acid promotes expression of germline-specific genes in chicken blastoderm cells by stimulating Smad1/5 phosphorylation in a feeder-free culture system

机译：维甲酸通过刺激无饲养层培养系统中的Smad1 / 5磷酸化来促进鸡胚细胞中种系特异性基因的表达

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BackgroundProducing transgenic chickens with chicken blastodermal cells (cBCs) is inefficient due to the extremely low germline transmission capacity of cBCs. As chicken primordial germ cells (PGCs) have been reported as an efficient method for producing transgenic chickens, the inefficiency of cBCs could potentially be resolved by inducing them to differentiate into germ cells. However, whether chemical inducers are able to enhance cBCs germline competence in vitro is unknown and the molecular mechanisms of differentiation of chicken pluripotent cells into germ cells are poorly understood. ResultsWe cultured cBCs with a monolayer morphology in E8 medium, a xeno- and feeder-free medium. We showed that retinoic acid (RA) treatment increased expression of germ cell-specific genes in cBCs. Using western blot, we determined that RA stimulated Smad1/5 phosphorylation. Moreover, Smad1/5 activation regulates the expression of germ cell-specific genes, as co-treatment with a Smad1/5 phosphorylation inhibitor or activator alters expression of these genes. We also demonstrate that Smad1/5 is required for RA-induced differentiation by RNA interference knockdown. ConclusionOur results demonstrated that E8 medium is able to maintain cBC growth for weeks and RA treatment induced germ cell differentiation of cBCs through the BMP-Smad1/5 signaling pathway.
机译：背景技术由于鸡胚细胞（cBC）的转基因鸡的种系传递能力极低，因此生产效率低下。由于已经报道了鸡原始生殖细胞（PGC）是生产转基因鸡的有效方法，因此通过诱导它们分化为生殖细胞可以潜在地解决cBC的低效率问题。然而，化学诱导剂是否能够在体外增强cBCs的种系能力尚不清楚，并且对鸡多能细胞分化为生殖细胞的分子机制了解甚少。结果我们在无异种和无饲养层的E8培养基中培养了具有单层形态的cBC。我们显示，视黄酸（RA）处理可增加cBC中生殖细胞特异性基因的表达。使用蛋白质印迹，我们确定RA刺激了Smad1 / 5磷酸化。此外，Smad1 / 5激活调节生殖细胞特异性基因的表达，因为与Smad1 / 5磷酸化抑制剂或激活剂共同处理可改变这些基因的表达。我们还证明Smad1 / 5是RNA干扰敲低RA诱导分化所必需的。结论我们的结果表明，E8培养基能够维持cBC生长数周，并且RA处理通过BMP-Smad1 / 5信号通路诱导cBC的生殖细胞分化。

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