Presumptive germ cells derived from mouse pluripotent somatic cell hybrids
Presumptive germ cells derived from mouse pluripotent somatic cell hybrids
复制标题
DOI：
10.1016/j.diff.2009.07.003
复制
发表时间：
2009-09-01
期刊：
影响因子：
2.9
通讯作者：
Kerkis, Irina
中科院分区：
文献类型：
--
作者：
Lavagnolli, Thais M. C.;Fonseca, Simone A. S.;Kerkis, Irina
胚胎干（ES）细胞与分化的体细胞融合是一种将体细胞核逆转为多能性状态的方法。由此产生的 ES-体细胞杂种 (ES-SCH) 保留了 ES 细胞的大部分特性：分化为多种细胞类型，并具有产生拟胚体 (EB) 和嵌合体的能力。然而，ES-SCH是否能够像ES细胞一样在体外完成分化为生殖细胞（GC）仍是未知数。在这里，我们表明，通过小鼠 ES 和脾细胞融合获得的近二倍体 ES-SCH 能够在体外分化为假定的 GC。 ES-SCH 的分化是通过 EB 形成和添加视黄酸诱导的。获得的推定GC与抗EMA、Vasa、Fragilis和Dazl抗体呈阳性反应，并表达GC特异性基因，例如Vasa、Stella、Dazl、Piwii 2、Tex 14、Bmp8b、Tdrd 1和Rnf17。荧光原位杂交分析表明 GC 样细胞中染色体减少。还检测到减数分裂和减数分裂后 GC 特异性基因（例如 Haprin、Acrosin、Scyp1、Scyp3 和 Stra-8）的表达。透射电子显微镜证实 ES-SCH 分化为推定 GC。来自“体细胞”伴侣的几个常染色体和 X 染色体的存在并不能阻止 ES-SCH 分化为假定的 GC。总体而言，我们的研究提出了一个有趣的体外模型，该模型允许研究重编程体细胞中的 GC 分化。(C) 2009 年国际分化学会。由 Elsevier Ltd 出版。保留所有权利。
The fusion of embryonic stem (ES) cells with differentiated somatic cells is an approach that reverses a somatic cell nucleus to a state of pluripotency. The resulting ES-somatic cell hybrids (ES-SCH) retain most of the properties of ES cells: differentiate into multiple cell types and have the ability to produce embryoid bodies (EB) and chimeras. However, it is still unknown whether ES-SCH will be able to complete the differentiation into germ cells (GC) in vitro similar to ES cells. Here, we show that near diploid ES-SCH, obtained by the fusion of mouse ES and spleen cells, were able to differentiate in vitro into presumptive GC. Differentiation of ES-SCH was induced through EB formation and by the addition of retinoic acid. Presumptive GC obtained reacted positively with anti-EMA, Vasa, Fragilis and Dazl antibodies and expressed GC-specific genes, such as Vasa, Stella, Dazl, Piwii 2, Tex 14, Bmp8b, Tdrd 1 and Rnf17. Fluorescent in situ hybridization analysis indicates chromosome reduction in the GC-like cells. Expression of meiotic and postmeiotic GC-specific genes such as Haprin, Acrosin, Scyp1, Scyp3 and Stra-8 were also detected. Transmission electron microscopy confirmed ES-SCH differentiation into presumptive GC. The presence of several autosomes and the X chromosome originated from the "somatic'' partner did not prevent ES-SCH differentiation towards presumptive GC. Overall our study suggests an interesting in vitro model, which allows the study GC differentiation in reprogrammed somatic cells. (C) 2009 International Society of Differentiation. Published by Elsevier Ltd. All rights reserved.