title: Stemness of spermatogonial stem cells encapsulated in alginate hydrogel during cryopreservation.
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 (6 days 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.
authors.affiliation: Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: A
authors.initials: A
authors.lastname: Pirnia
authors.affiliation: Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: K
authors.initials: K
authors.lastname: Parivar
authors.affiliation: Fertility and Infertility Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
authors.forename: M
authors.initials: M
authors.lastname: Hemadi
authors.affiliation: Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: P
authors.initials: P
authors.lastname: Yaghmaei
authors.affiliation: Razi Herbal Medicine Research center and department of Anatomical sciences, Lorestan University of Medical Sciences, Khorramabad, Iran.
authors.forename: M
authors.initials: M
authors.lastname: Gholami
mesh_terms: D000464:Alginates; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D004268:DNA-Binding Proteins / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D007248:Infertility, Male; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016601:RNA-Binding Proteins / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D013091:Spermatogenesis; D013093:Spermatogonia / Q000166:cytology* / Q000502:physiology; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000502:physiology*; D013737:Testis / Q000166:cytology; D014157:Transcription Factors / Q000235:genetics
chemical_list: D000464:Alginates; D003451:Cryoprotective Agents; D004268:DNA-Binding Proteins; D006603:Hexuronic Acids; C488352:Lin-28 protein, mouse; D016601:RNA-Binding Proteins; C509167:Sall4 protein, mouse; D014157:Transcription Factors; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D020723:Glucuronic Acid
keywords: alginate hydrogel; cryopreservation; spermatogonial stem cells; stemness genes