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Evaluation of co-cultured spermatogonial stem cells encapsulated in alginate hydrogel with Sertoli cells and their transplantation into azoospermic mice

Published online by Cambridge University Press:  06 October 2021

Mohammad Veisi ,
Kamran Mansouri ,
Vahideh Assadollahi ,
Cyrus Jalili ,
Afshin Pirnia ,
Mohammad Reza Salahshoor ,
Zohreh Hoseinkhani  and
Mohammad Reza Gholami Open the ORCID record for Mohammad Reza Gholami [Opens in a new window]
Mohammad Veisi
Affiliation:
Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
Kamran Mansouri
Affiliation:
Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
Vahideh Assadollahi
Affiliation:
Medical Technology Research Center, Institute Health Technology Kermanshah University of Medical Sciences, Kermanshah, Iran
Cyrus Jalili
Affiliation:
Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
Afshin Pirnia
Affiliation:
Medical Technology Research Center, Institute Health Technology Kermanshah University of Medical Sciences, Kermanshah, Iran
Mohammad Reza Salahshoor
Affiliation:
Medical Technology Research Center, Institute Health Technology Kermanshah University of Medical Sciences, Kermanshah, Iran
Zohreh Hoseinkhani
Affiliation:
Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
Mohammad Reza Gholami*
Affiliation:
Medical Technology Research Center, Institute Health Technology Kermanshah University of Medical Sciences, Kermanshah, Iran
*
Author for correspondence: Mohammadreza Gholami. Medical Technology Research Center, Institute Health Technology Kermanshah University of Medical Sciences, Kermanshah, Iran. E-mail: rezagholami57@gmail.com
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Summary

An in vitro spermatogonial stem cell (SSC) culture can serve as an effective technique to study spermatogenesis and treatment for male infertility. In this research, we compared the effect of a three-dimensional alginate hydrogel with Sertoli cells in a 3D culture and co-cultured Sertoli cells. After harvest of SSCs from neonatal mice testes, the SSCs were divided into two groups: SSCs on a 3D alginate hydrogel with Sertoli cells and a co-culture of SSCs with Sertoli cells for 1 month. The samples were evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays and bromodeoxyuridine (BrdU) tracing, haematoxylin and eosin (H&E) and periodic acid–Schiff (PAS) staining after transplantation into an azoospermic testis mouse. The 3D group showed rapid cell proliferation and numerous colonies compared with the co-culture group. Molecular assessment showed significantly increased integrin alpha-6, integrin beta-1, Nanog, Plzf, Thy-1, Oct4 and Bcl2 expression levels in the 3D group and decreased expression levels of P53, Fas, and Bax. BrdU tracing, and H&E and PAS staining results indicated that the hydrogel alginate improved spermatogenesis after transplantation in vivo. This finding suggested that cultivation of SSCs on alginate hydrogel with Sertoli cells in a 3D culture can lead to efficient proliferation and maintenance of SSC stemness and enhance the efficiency of SSC transplantation.

Keywords

Alginate hydrogelApoptosisCo-cultureSertoliSpermatogonial stem cellsStemness genesTransplantation
Type
Research Article
Information
Zygote , Volume 30 , Issue 3 , June 2022 , pp. 344 - 351
Copyright
© The Author(s), 2021. Published by Cambridge University Press

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