Stemness of spermatogonial stem cells encapsulated in alginate hydrogel during cryopreservation

Pirnia, A and Parivar, K and Hemadi, M and Yaghmaei, P and Gholami, M (2017) Stemness of spermatogonial stem cells encapsulated in alginate hydrogel during cryopreservation. ANDROLOGIA, 49. ISSN 0303-4569

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Abstract

This study investigated the effect of spermatogonial stem cell encapsulated in alginate hydrogel during cryopreservation, as cells were protected against damage during cryopreservation within the hydrogel. Spermatogonial stem cells were isolated from the testes of Balb/c mice pups (6days old), purified in laminin-coated dishes and CD90.1 microbeads, encapsulated in alginate hydrogel and then cryopreserved. After thawing, cell viability and Spermatogonial stem cell (SSC) colony diameter were evaluated. After RNA was isolated and cDNA was synthesised, the expression of stemness genes was considered using RT real-time PCR. Finally, spermatogonial stem cells labelled with BrdU were transplanted to busulfan azoospermic mouse models. Lin28a and Sall4 genes were significantly upregulated after cryopreservation in alginate hydrogel. However, cell viability was significantly decreased. The diameter of colonies consisting of spermatogonial stem cells freeze-thawed in alginate microbeads showed no significant difference with fresh spermatogonial stem cells and the control group. The injection of freeze-thawed spermatogonial stem cells encapsulated in alginate hydrogel resulted in spermatogenesis recovery. Alginate mimics the extracellular matrices (ECM) for spermatogonial stem cells; therefore, it can support stemness potential during the cell cryopreservation process and restart spermatogenesis after transplantation. Keywords Author Keywords:alginate hydrogel; cryopreservation; spermatogonial stem cells; stemness genes KeyWords Plus:TESTICULAR TISSUE; GERM-CELLS; FERTILITY PRESERVATION; COLONY FORMATION; SELF-RENEWAL; IN-VITRO; SEMINIFEROUS TUBULES; A SPERMATOGONIA; STROMAL CELLS; MOUSE TESTIS

Item Type: Article
Subjects: R Medicine > R Medicine (General)
Divisions: Faculty of Medicine, Health and Life Sciences > School of Medicine
Depositing User: sobhan rezaiian
Date Deposited: 07 Dec 2017 18:59
Last Modified: 07 Dec 2017 18:59
URI: http://eprints.lums.ac.ir/id/eprint/1025

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