Characterisation and germline transmission of cultured avian primordial germ cells.
Characterisation and germline transmission of cultured avian primordial germ cells.
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培养的禽类原始生殖细胞的特性分析与种系传递
DOI：
10.1371/journal.pone.0015518
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发表时间：
2010-11-29
期刊：
影响因子：
3.7
通讯作者：
McGrew MJ
中科院分区：
文献类型：
--
作者：
Macdonald J;Glover JD;Taylor L;Sang HM;McGrew MJ
鸟类原始生殖细胞（Primordial Germ Cells，PGCs）作为一种基于细胞的系统，在鸟类种质资源的研究和保存以及基因组的遗传修饰方面具有重要的应用前景。以前有报道说，鸡胚胎的PGCs可以在培养物中繁殖，并有助于宿主鸟类的生殖细胞谱系。我们证实了这些结果表明，PGCs从不同的层品种的鸡可以在体外繁殖延长的时间。我们证明，通过PI 3 K和MEK的细胞内信号传导对于PGC生长是必要的。我们对这些细胞进行了初步鉴定。我们发现培养的PGCs含有大的脂质空泡，富含糖原，并表达干细胞标志物SSEA-1。这些细胞还表达生殖细胞特异性蛋白CVH和CDH。出乎意料的是，使用RT-PCR，我们显示培养的PGCs表达多能性基因c-Myc、cKlf 4、cPouV、cSox 2和cNanog。最后，我们证明了培养的PGCs将迁移到宿主胚胎的形成性腺并定殖。雄性PGCs将定殖于雌性性腺并进入减数分裂，但在性发育期间从性腺中丢失。在雄性宿主中，培养的PGCs形成有功能的配子，如通过产生存活后代所证明的。胚系感受态禽类PGCs体外培养的建立为研究早期生殖细胞分化提供了一个独特的体系，也为哺乳动物生殖细胞发育提供了一个比较体系。初级PGC系将形成用于保存禽类种质的替代技术的基础，并且将是转基因技术的有价值的工具，具有研究和工业应用。
Avian primordial germ cells (PGCs) have significant potential to be used as a cell-based system for the study and preservation of avian germplasm, and the genetic modification of the avian genome. It was previously reported that PGCs from chicken embryos can be propagated in culture and contribute to the germ cell lineage of host birds. We confirm these results by demonstrating that PGCs from a different layer breed of chickens can be propagated for extended periods in vitro. We demonstrate that intracellular signalling through PI3K and MEK is necessary for PGC growth. We carried out an initial characterisation of these cells. We find that cultured PGCs contain large lipid vacuoles, are glycogen rich, and express the stem cell marker, SSEA-1. These cells also express the germ cell-specific proteins CVH and CDH. Unexpectedly, using RT-PCR we show that cultured PGCs express the pluripotency genes c-Myc, cKlf4, cPouV, cSox2, and cNanog. Finally, we demonstrate that the cultured PGCs will migrate to and colonise the forming gonad of host embryos. Male PGCs will colonise the female gonad and enter meiosis, but are lost from the gonad during sexual development. In male hosts, cultured PGCs form functional gametes as demonstrated by the generation of viable offspring. The establishment of in vitro cultures of germline competent avian PGCs offers a unique system for the study of early germ cell differentiation and also a comparative system for mammalian germ cell development. Primary PGC lines will form the basis of an alternative technique for the preservation of avian germplasm and will be a valuable tool for transgenic technology, with both research and industrial applications.
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影响因子：
64.5
作者：
HAY, B;JAN, LY;JAN, YN
通讯作者：
JAN, YN
影响因子：
2.7
作者：
Farini, D;Scaldaferri, ML;De Felici, M
通讯作者：
De Felici, M
影响因子：
5.2
作者：
Jung, JS;Kim, DK;Han, JY
通讯作者：
Han, JY
影响因子：
64.8
作者：
HENRIQUE, D;ADAM, J;ISHHOROWICZ, D
通讯作者：
ISHHOROWICZ, D
DOI：
10.1098/rspb.1975.0086
发表时间：
1975-01-01
期刊：
PROCEEDINGS OF THE ROYAL SOCIETY SERIES B-BIOLOGICAL SCIENCES
影响因子：
--
作者：
FORD, CE;EVANS, EP;BARNES, RD
通讯作者：
BARNES, RD