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      Stemness of spermatogonial stem cells encapsulated in alginate hydrogel during cryopreservation.

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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 (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.

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          Author and article information

          Journal
          Andrologia
          Andrologia
          Wiley-Blackwell
          1439-0272
          0303-4569
          Jun 2017
          : 49
          : 5
          Affiliations
          [1 ] Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
          [2 ] Fertility and Infertility Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
          [3 ] Razi Herbal Medicine Research center and department of Anatomical sciences, Lorestan University of Medical Sciences, Khorramabad, Iran.
          Article
          10.1111/and.12650
          27469285
          e2aab52a-a31a-4b34-a3bf-2e844d0e4906
          History

          alginate hydrogel,cryopreservation,spermatogonial stem cells,stemness genes

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