Pluripotency Potential of Embryonic Stem Cell-Like Cells Derived From Mouse Testis

Objective: during the cultivation of spermatogonial stem cells (sscs) and their conversion into embryonic stem-like (es-like) cells, transitional es-like colonies and epiblast-like cells were observable. in the present experimental study, we aimed to analyze the efficiency of the multipotency or pluripotency potential of es-like cells, transitional colonies and epiblast-like cells. materials and methods: in this experimental study, sscs were isolated from transgenic octamer-binding transcription factor 4 (oct4)-green fluorescent protein (gfp)-reporter mice. during cell culture es-like, transitional and epiblastlike colonies developed spontaneously. the mrna and protein expression of pluripotency markers were analyzed by fluidigm real-time polymerase chain reaction (rt-pcr) and immunocytochemistry, respectively. efficiency to produce chimera mice was evaluated after injection of es and es-like cells into blastocysts. results: microscopic analyses demonstrated that the expression of oct4-gfp in es-like cells was very strong, in epiblast-like cells was not detectable, and was only partial in transitional colonies. fluidigm rt-pcr showed a higher expression of the germ cell markers stra-8 and gpr-125 in es-like cells and the pluripotency genes dppa5, lin28, klf4, gdf3 and tdgf1 in es-like colonies and embryonic stem cells (escs) compared to the epiblast-like and transitional colonies. no significant expression of oct-4, nanog, sox2 and c-myc was observed in the different groups. we showed a high expression level of nanog and klf4 in es-like, while only a partial expression was observed in transitional colonies. we generated chimeric mice after blastocystic injection from es and es-like cells, but not from transitional colonies. we observed that the efficiency to produce chimeric mice in es cells was more efficient (59%) in comparison to es-like cells (22%). conclusion: this new data provides more information on the pluripotency or multipotency potentials of testis-derived es-like cells in comparison to transitional colonies and epiblast-like cells.