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Iran Red Crescent Med J. 2012 Dec;14(12):811-5. doi: 10.5812/ircmj.4051. Epub 2012 Dec 6.

A Simple Co-culture System for Generation of Embryonic Stem-Like Cells From Testis.

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1
Department of Anatomical Sciences, Medical Sciences Faculty, Tarbiat Modares University, Tehran, IR Iran.

Abstract

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.

KEYWORDS:

Reprogramming; Spermatogonia; Testis

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