Pluripotent cell derivation from male germline cells by suppression of <i>Dmrt1</i> and <i>Trp53</i>
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Diploid germ cells are thought to have pluripotency potential. We recently described a method to derive pluripotent stem cells (PSCs) from cultured spermatogonial stem cells (SSCs) by depleting <i>Trp53</i> and <i>Dmrt1</i>, both of which are known suppressors of teratomas. In this study, we used this technique to analyze the effect of this protocol in deriving PSCs from the male germline at different developmental stages. We collected primordial germ cells (PGCs), gonocytes and spermatogonia, and the cells were transduced with lentiviruses expressing short hairpin RNA against <i>Dmrt1</i> and/or <i>Trp53</i>. We found that PGCs are highly susceptible to reprogramming induction and that only <i>Trp53</i> depletion was sufficient to induce pluripotency. In contrast, gonocytes and spermatogonia were resistant to reprogramming by double knockdown of <i>Dmrt1</i> and <i>Trp53</i>. PSCs derived from PGCs contributed to chimeras produced by blastocyst injection, but some of the embryos showed placenta-only phenotypes suggestive of epigenetic abnormalities of PGC-derived PSCs. These results show that PGCs and gonocytes/spermatogonia have distinct reprogramming potential and also suggest that fresh and cultured SSCs do not necessarily have the same properties.
- Journal of Reproduction and Development
Journal of Reproduction and Development 61(5), 473-484, 2015
Japanese Society of Animal Reproduction (JSAR)