Derivation of primordial germ cells from human embryonic and induced pluripotent stem cells is significantly improved by coculture with human fetal gonadal cells.
Derivation of primordial germ cells from human embryonic and induced pluripotent stem cells is significantly improved by coculture with human fetal gonadal cells.
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DOI：
10.1002/stem.13
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发表时间：
2009-04
期刊：
影响因子：
5.2
通讯作者：
Clark, Amander T.
中科院分区：
文献类型：
--
作者：
Park, Tae Sub;Galic, Zoran;Conway, Anne E.;Lindgren, Anne;van Handel, Benjamin J.;Magnusson, Mattias;Richter, Laura;Teitell, Michael A.;Mikkola, Hanna K. A.;Lowry, William E.;Plath, Kathrin;Clark, Amander T.
从人胚胎干细胞（hESC）或人诱导多能干细胞（hIPS）衍生生殖细胞代表了治疗不孕症的理想实验模型和潜在策略。在目前的研究中，我们开发了一种三重生物标志物检测方法，用于识别和分离人原始生殖细胞（PGC），首先评估体内前三个月期间人PGC的形成。接下来，我们将该技术应用于表征来自多能细胞的体外衍生的PGCs（iPGCs）。我们的结果表明，hESC在人胎儿性腺基质细胞上的共分化显著提高了产生iPGC的效率。此外，效率在各种多能细胞系之间是相当的，而不管来源于人胚泡的内细胞团（hESC）或人皮肤成纤维细胞的重编程（hIPS）。为了更好地表征iPGC，我们进行了真实的时间PCR、微阵列和亚硫酸氢盐测序。我们的结果表明，iPGC在分化第7天在转录上与体细胞不同，表达与多能性和生殖细胞发育相关的基因，同时抑制与体细胞分化相关的基因（特别是多个HOX基因）。使用亚硫酸氢盐测序，我们发现iPGC在分化的第7天从差异甲基化的印迹区域开始印迹擦除。然而，来源于hIPS细胞的iPGC不能有效地启动印记擦除。总之，我们的结果表明，来源于在hFGS细胞上分化的多能细胞的三重阳性iPGC对应于已经启动印记擦除过程的定向的妊娠早期生殖细胞（9周之前）。
The derivation of germ cells from human embryonic stem cells (hESCs) or human induced pluripotent stem (hIPS) cells represents a desirable experimental model and potential strategy for treating infertility. In the current study we developed a triple biomarker assay for identifying and isolating human primordial germ cells (PGCs) by first evaluating human PGC formation during the first trimester in vivo. Next, we applied this technology to characterizing in vitro derived PGCs (iPGCs) from pluripotent cells. Our results show that co-differentiation of hESCs on human fetal gonadal stromal cells significantly improves the efficiency of generating iPGCs. Furthermore, the efficiency was comparable between various pluripotent cell lines regardless of origin from the inner cell mass of human blastocysts (hESCs), or reprogramming of human skin fibroblasts (hIPS). In order to better characterize the iPGCs we performed Real time PCR, microarray and bisulfite sequencing. Our results show that iPGCs at day 7 of differentiation are transcriptionally distinct from the somatic cells, expressing genes associated with pluripotency and germ cell development while repressing genes associated with somatic differentiation (specifically multiple HOX genes). Using bisulfite sequencing, we show that iPGCs initiate imprint erasure from differentially methylated imprinted regions by day 7 of differentiation. However, iPGCs derived from hIPS cells do not initiate imprint erasure as efficiently. In conclusion, our results indicate that triple positive iPGCs derived from pluripotent cells differentiated on hFGS cells correspond to committed first trimester germ cells (before 9 weeks) that have initiated the process of imprint erasure.
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影响因子：
64.8
作者：
Hajkova, Petra;Ancelin, Katia;Waldmann, Tanja;Lacoste, Nicolas;Lange, Ulrike C.;Cesari, Francesca;Lee, Caroline;Almouzni, Genevieve;Schneider, Robert;Surani, M. Azim
通讯作者：
Surani, M. Azim
影响因子：
5.2
作者：
Kerr, Candace L.;Hill, Christine M.;Gearhart, John D.
通讯作者：
Gearhart, John D.
影响因子：
5.2
作者：
Bucay, Nathan;Yebra, Mayra;Montgomery, Anthony M. P.
通讯作者：
Montgomery, Anthony M. P.
DOI：
10.1073/pnas.241508898
发表时间：
2001-11-20
影响因子：
11.1
作者：
Fujiwara, T;Dunn, NR;Hogan, BLM
通讯作者：
Hogan, BLM
影响因子：
6.1
作者：
Chen, H. -F.;Kuo, H. -C.;Ho, H-N.
通讯作者：
Ho, H-N.