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Journal of Assisted Reproduction and Genetics

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01-09-2012 | Stem Cell Biology

Proliferation of small number of human spermatogonial stem cells obtained from azoospermic patients

Authors: Morteza Koruji, Abdulhossein Shahverdi, Arghavan Janan, Abbas Piryaei, Mohammad Reza Lakpour, Mohammad Ali Gilani Sedighi

Published in: Journal of Assisted Reproduction and Genetics | Issue 9/2012

Abstract

Purpose

This study aims to proliferate spermatogonial stem cells (SSCs) and compare the in-vitro effects of laminin and growth factors on the proliferation of adult human SSC.

Methods

Isolated testicular cells were cultured in DMEM supplemented with 5 % fetal calf serum (FCS). During the culture, enriched spermatogonial cells were treated with a combination of glial cell line-derived neurotrophic factor (GDNF), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) and mouse leukemia inhibitory factor (LIF) in the presence or absence of human placental laminin-coated dishes. Cluster assay was performed during culture. Presence of spermatogonia was determined by an ultrastructural study of the cell clusters, reverse transcription polymerase chain reaction (RT-PCR) for spermatogonial markers and xenotransplantation to the testes of busulfan-treated recipient mice. Statistical significance between mean values was determined using statistical ANOVA tests.

Results

The findings indicated that the addition of GDNF, bFGF, EGF and LIF on laminin-coated dishes significantly increased in-vitro spermatogonial cell cluster formation in comparison with the control group (p ≤ 0.001). The expression of spermatogonial markers was maintained throughout the culture period. Furthermore, a transplantation experiment showed the presence of SSC among the cultured cells. In addition, a transmission electron microscopy (TEM) study suggested the presence of spermatogonial cells of typical morphology among the cluster cells.

Conclusions

It can be concluded that human SSCs obtained from non-obstructive azoospermic (NOA) patients had the ability to self-renew in the culture system. This system can be used for the propagation of a small number of these cells from small biopsies.

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Title: Proliferation of small number of human spermatogonial stem cells obtained from azoospermic patients
Authors: Morteza Koruji
Abdulhossein Shahverdi
Arghavan Janan
Abbas Piryaei
Mohammad Reza Lakpour
Mohammad Ali Gilani Sedighi
Publication date: 01-09-2012
Publisher: Springer US
Published in: Journal of Assisted Reproduction and Genetics / Issue 9/2012
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI: https://doi.org/10.1007/s10815-012-9817-8

At a glance: The STEP trials

Springer Medicine

Proliferation of small number of human spermatogonial stem cells obtained from azoospermic patients

Abstract

Purpose

Methods

Results

Conclusions

At a glance: The STEP trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

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