Abstract
BackgroundNew 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.ObjectivesThe aim of the present study was to generate embryonic stem-like cells from neonatal mouse testis.Materials and MethodsThe 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.ResultsThe 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.ConclusionsThis researh indicates pluripotency evidence of ES-like cells derived from testis. ES-like cells shows some molecular characteristics with embryonic stem cells.
Highlights
New research proposes the pluripotency of spermatogonial cells obtained from testis
Pluripotent stem cells are characterized by their extensive proliferation and self-renewal ability and they can differentiate into many different cell types, this property makes them a renewable source which are usable for the treatment of disease and developmental research [1, 2].Different type of pluripotent stem cell is recognized in mammals
Embryonal carcinoma (EC) cells are derived from testicular tumors [3, 4].The recent research shows that embryonic stem-like cells can be generated from mouse spermatogonial stem cells (SSCs) under culture condition
Summary
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. 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; Stra, mvh, and piwill became downregulated. SSCs have the ability of spermatogenesis which produces gamet, in addition these cells can transit to pluripotent cells which differentiate into all three germ layers [6] They will be usable for basic research and studying the differentiation mechanisms of stem cell. They offer a new stem cells source for treatment of disease in human regenerative medicine [7] Some criteria, such as differentiation into three germ layers and contribution to chimeras, demonstrated the pluripotency of mouse
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