Pluripotency Potential of Embryonic Stem CellLike Cells Derived from Mouse Testis

Objective During the cultivation of spermatogonial stem cells (SSCs) and their conversion into embryonic stemlike (ESlike) cells, transitional ESlike colonies and epiblastlike cells were observable. In the present experimental study, we aimed to analyze the efficiency of the multipotency or pluripotency potential of ESlike cells, transitional colonies and epiblastlike cells. Materials and Methods In this experimental study, SSCs were isolated from transgenic octamerbinding transcription factor 4 (Oct4)green fluorescent protein (GFP)reporter mice. During cell culture ESlike, transitional and epiblast like colonies developed spontaneously. The mRNA and protein expression of pluripotency markers were analyzed by Fluidigm realtime polymerase chain reaction (RTPCR) and immunocytochemistry, respectively. Efficiency to produce chimera mice was evaluated after injection of ES and ESlike cells into blastocysts. Results Microscopic analyses demonstrated that the expression of Oct4GFP in ESlike cells was very strong, in epiblastlike cells was not detectable, and was only partial in transitional colonies. Fluidigm RTPCR showed a higher expression of the germ cell markers Stra8 and Gpr125 in ESlike cells and the pluripotency genes Dppa5, Lin28, Klf4, Gdf3 and Tdgf1 in ESlike colonies and embryonic stem cells (ESCs) compared to the epiblastlike and transitional colonies. No significant expression of Oct4, Nanog, Sox2 and cMyc was observed in the different groups. We showed a high expression level of Nanog and Klf4 in ESlike, while only a partial expression was observed in transitional colonies. We generated chimeric mice after blastocystic injection from ES and ESlike 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 ESlike cells (22%). ConclusionThis new data provides more information on the pluripotency or multipotency potentials of testisderived ESlike cells in comparison to transitional colonies and epiblastlike cells.