Fate of induced pluripotent stem cells following transplantation to murine seminiferous tubules

被引:45
作者
Durruthy, Jens Durruthy [1 ,2 ]
Ramathal, Cyril [1 ,2 ]
Sukhwani, Meena [3 ]
Fang, Fang [1 ,2 ]
Cui, Jun [1 ,2 ]
Orwig, Kyle E. [3 ]
Pera, Renee A. Reijo [1 ,2 ]
机构
[1] Stanford Univ, Dept Genet, Stanford, CA 94305 USA
[2] Stanford Univ, Dept Obstet & Gynecol, Inst Stem Cell Biol & Regenerat Med, Ctr Reprod & Stem Cell Biol, Stanford, CA 94305 USA
[3] Univ Pittsburgh, Sch Med, Magee Womens Res Inst, Dept Obstet Gynecol & Reprod Sci, Pittsburgh, PA 15213 USA
基金
美国国家卫生研究院;
关键词
PRIMORDIAL GERM-CELLS; IN-VITRO; GENE-EXPRESSION; IPS CELLS; DIFFERENTIATION; VASA; 5-HYDROXYMETHYLCYTOSINE; BIOLOGY; FETAL; DAZL;
D O I
10.1093/hmg/ddu012
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:3071 / 3084
页数:14
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