Hybridization of testis-derived stem cells with somatic cells and embryonic stem cells in Mice
Hybridization of testis-derived stem cells with somatic cells and embryonic stem cells in Mice
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小鼠睾丸干细胞与体细胞和胚胎干细胞的杂交
DOI：
10.1095/biolreprod.112.098988
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
T
中科院分区：
文献类型：
--
作者：
Takehashi;M.;Tada;M.;Kanatsu-Shinohara;M.;Morimoto;H.;Kazuki;Y.;Oshimura;M.;Tada;T. and Shinohara;T
体细胞杂交被广泛用于研究基因调控的控制和分化状态的稳定性。相比之下，这种方法在生殖细胞上的应用受到限制，部分原因是无法培养生殖细胞。在本研究中，我们利用生殖系干细胞（GS）和多能生殖系干细胞（mGS）制备了生殖细胞杂种。虽然GS细胞具有精原干细胞（SSC）活性，但mGS细胞与胚胎干细胞（ES）细胞相似，最初来源于GS细胞。成功地获得了GS细胞与ES细胞、GS细胞与mGS细胞、mGS细胞与胸腺细胞的杂交。所有小鼠均表现出胚胎干细胞标记物和与胚胎干细胞相似的行为，形成畸胎瘤，并分化为体细胞组织。然而，将这些杂交细胞显微注射到精管后，没有一个细胞能够重建精子发生。对印迹基因的DNA甲基化模式的分析也表明，mGS细胞不具有DNA去甲基化能力，这在源自原始生殖细胞的胚胎生殖细胞中发现。然而，mGS细胞以类似于ES细胞的方式重新激活X染色体并诱导雌性胸腺细胞中的pou5f1表达。这些数据表明，mGS细胞具有es样重编程潜力，这种潜力主导着过ssc活性。
Somatic cell hybridization is widely used to study the control of gene regulation and the stability of differentiated states. In contrast, the application of this method to germ cells has been limited in part because of an inability to culture germ cells. In this study, we produced germ cell hybrids using germ-line stem (GS) cells and multipotent germ-line stem (mGS) cells. While GS cells are enriched for spermatogonial stem cell (SSC) activity, mGS cells are similar to embryonic stem (ES) cells and originally derived from GS cells. Hybrids were successfully obtained between GS cells and ES cells, between GS cells and mGS cells, and between mGS cells and thymocytes. All exhibited ES cell markers and a behavior similar to ES cells, formed teratomas, and differentiated into somatic cell tissues. However, none of the hybrid cells were able to reconstitute spermatogenesis after microinjection into seminiferous tubules. Analyses of the DNA methylation patterns of imprinted genes also showed that mGS cells do not possess a DNA demethylation ability, which was found in embryonic germ cells derived from primordial germ cells. However, mGS cells reactivated the X chromosome and inducedPou5f1expression in female thymocytes in a manner similar to ES cells. These data show that mGS cells possess ES-like reprogramming potential, which predominates over-SSC activity.