Induction of Pluripotency in Adult Unipotent Germline Stem Cells

Mouse and human stem cells with features similar to those of embryonic stem cells have been derived from testicular cells. Although pluripotent stem cells have been obtained from defined germline stem cells (GSCs) of mouse neonatal testis, only multipotent stem cells have been obtained so far from defined cells of mouse adult testis. In this study we describe a robust and reproducible protocol for obtaining germline-derived pluripotent stem (gPS) cells from adult unipotent GSCs. Pluripotency of gPS cells was confirmed by in vitro and in vivo differentiation, including germ cell contribution and transmission. As determined by clonal analyses, gPS cells indeed originate from unipotent GSCs. We propose that the conversion process requires a GSC culture microenvironment that depends on the initial number of plated GSCs and the length of culture time. 20 samples in total. Germline stem cells (GSC): - GSC_1 - GSC_2 - GSC_3 Germline pluripotent stem cells (gPS): - gPSC_1 - gPSC_2 - gPSC_3 - gPSC_4 - gPSC_5 Neural stem cells (NSC): - NSC_1 - NSC_2 - NSC_3 Mouse embryonic fibroblast (MEF): - MEF_1 - MEF_2 - MEF_3 Embryonic stem cells (ESC): - ESC_1 - ESC_2 - ESC_3 ESC-like cells (ESC_like) - ESC_like_1 - ESC_like_2 - ESC_like_3