Fate of induced pluripotent stem cells following transplantation to murine seminiferous tubules.
Fate of induced pluripotent stem cells following transplantation to murine seminiferous tubules.
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DOI：
10.1093/hmg/ddu012
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发表时间：
2014-06-15
影响因子：
3.5
通讯作者：
Reijo Pera RA
中科院分区：
文献类型：
--
作者：
Durruthy Durruthy J;Ramathal C;Sukhwani M;Fang F;Cui J;Orwig KE;Reijo Pera RA
人类生殖细胞发育的研究在很大程度上受到生殖细胞发育过程中不可接近性的限制。此外，尽管有几项研究报道了小鼠和人类生殖细胞在体外从多能干细胞（PSCs）分化，但在体内从多能干细胞分化人类生殖细胞还没有报道。在这里，我们测试了mRNA重编程结合异种移植是否可以提供一个可行的系统，通过诱导多能干细胞（iPSCs）来探索人类生殖细胞发育的遗传学。为此，我们通过单独使用OCT3/4、SOX2、KLF4和cMYC （OSKM）或结合生殖细胞特异性mRNA VASA （OSKMV）进行基于mRNA的重编程，获得了无整合的iPSCs。所有iPSC系均符合多能性的经典标准。此外，全局基因表达谱没有区分未分化的OSKM和OSKMV iPSCs之间的大差异；然而，在多能因子和生殖细胞特异性基因的表达、表观遗传谱和体外分化研究中发现了一些差异。相比之下，将未分化的iPSCs直接移植到生殖细胞缺失免疫缺陷小鼠的精小管中，不同因素产生的iPSCs的命运不同。移植在体内产生形态学和免疫组织化学上可识别的生殖细胞，特别是OSKMV细胞。值得注意的是，OSKMV细胞也不形成肿瘤，而留在精小管外的OSKM细胞广泛增殖并形成肿瘤。结果表明，结合移植的mRNA重编程可以为人类生殖细胞发育的遗传分析提供工具。
Studies of human germ cell development are limited in large part by inaccessibility of germ cells during development. Moreover, although several studies have reported differentiation of mouse and human germ cells from pluripotent stem cells (PSCs) in vitro, differentiation of human germ cells from PSCs in vivo has not been reported. Here, we tested whether mRNA reprogramming in combination with xeno-transplantation may provide a viable system to probe the genetics of human germ cell development via use of induced pluripotent stem cells (iPSCs). For this purpose, we derived integration-free iPSCs via mRNA-based reprogramming with OCT3/4, SOX2, KLF4 and cMYC alone (OSKM) or in combination with the germ cell-specific mRNA, VASA (OSKMV). All iPSC lines met classic criteria of pluripotency. Moreover, global gene expression profiling did not distinguish large differences between undifferentiated OSKM and OSKMV iPSCs; however, some differences were observed in expression of pluripotency factors and germ cell-specific genes, and in epigenetic profiles and in vitro differentiation studies. In contrast, transplantation of undifferentiated iPSCs directly into the seminiferous tubules of germ cell-depleted immunodeficient mice revealed divergent fates of iPSCs produced with different factors. Transplantation resulted in morphologically and immunohistochemically recognizable germ cells in vivo, particularly in the case of OSKMV cells. Significantly, OSKMV cells also did not form tumors while OSKM cells that remained outside the seminiferous tubule proliferated extensively and formed tumors. Results indicate that mRNA reprogramming in combination with transplantation may contribute to tools for genetic analysis of human germ cell development.
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DOI：
10.1126/science.1229277
发表时间：
2013-01-25
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Hackett JA;Sengupta R;Zylicz JJ;Murakami K;Lee C;Down TA;Surani MA
通讯作者：
Surani MA
影响因子：
3.5
作者：
Kerjean, A;Dupont, JM;Jeanpierre, M
通讯作者：
Jeanpierre, M
影响因子：
4
作者：
Gustafson, Eric A.;Wessel, Gary M.
通讯作者：
Wessel, Gary M.
影响因子：
64.8
作者：
FERGUSONSMITH, AC;SASAKI, H;SURANI, MA
通讯作者：
SURANI, MA
影响因子：
15.9
作者：
Dovey, Serena L.;Valli, Hanna;Orwig, Kyle E.
通讯作者：
Orwig, Kyle E.