a simple co-culture system for generation of embryonic stem-like cells from testis

background new research proposes the pluripotency of spermatogonial cells obtained from testis. these spermatogonia-derived stem cells are called embryonic stem-like cells that express embryonic stem cell markers and differentiate to the three germ layers. objectives the aim of the present study was to generate embryonic stem-like cells from neonatal mouse testis. materials and methods the testis cells were collected from neonatal mouse. after decapsulation, testis was mechanically dissected and dissociated via a two-step mechanical and enzymatic digestion. the spermatogonia and sertoli cells were cultured together in dulbecco’s modified eagle’s medium (dmem) supplemented with 15% fbs and lif. before one week, several small spermatogonia colonies were observed on top of the monolayer of sertoli cells. these colonies were passaged every four days. es-like cells colonies that resembled es cell was appeared within 2-3 weeks (at passages 5). real time pcr was performed to analyze the expression of a subset of pluripotency markers, as well as germ cell-specific genes. es like cells were confirmed with ssea1, sox2 and oct4 immunofluorescence stainng as pluripotency markers. results the results showed that at fifth passages, the pluripotency genes; nanog and c-myc have significant increase in es-like cells in compare with spermatogonia cells, whereas the spermatogonial markers; stra8, mvh, and piwill2 became downregulated. in addition to these pluripotency genes, the es cell marker ssea-1, sox2 and oct4 were expressed in the es-like cells, similar to es cells. conclusions this researh indicates pluripotency evidence of es-like cells derived from testis. es-like cells shows some molecular characteristics with embryonic stem cells.