Study Effect of Encapsulation of Spermatogonial Stem Cells in Alginate Hydrogel During Cryopreservation
DOI:
https://doi.org/10.22100/jkh.v11i4.1550Keywords:
Spermatogonial Stem Cells, Cryopreservation, Alginate.Abstract
Introduction: Spermatogonial stem cells (SSCs) are the foundation of spermatogenesis. Cancer treatments such as chemotherapy or radiotherapy by targeting of these high proliferating cells can cause infertility. In pubertal male patients, cryopreservation of spermatozoa before chemotherapy is the way for fertility preservation during treatment. However, for prepubertal male patients, because of the absence of spermatozoids, this is not possible. The aim of this study is the investigation of the effect of alginate hydrogel in reduction of cryopreservation effects on SSCs potential and preserve of these cells in cancer childhoods that undergoes chemotherapy or radiotherapy.
Methods: SSCs were isolated from the testes of Balb/c mice pups (6 days old), purified by MACS with Thy-1 and c-kit microbeads, encapsulated in alginate hydrogel and then cryopreserved. After thawing, cell viability was evaluated. After RNA was isolated and cDNA was synthesized, the expression of stemness genes Oct4, Sall4, Plzf, Dazl, Etv5, Bcl6b, Lin28, and Nanog was considered using RT Real-time PCR. Statistical analyses were performed using SPSS. Gene expression was analyzed with REST software.
Results: Cell viability after cryopreservation show no significant decrease in alginate encapsulated cells group compared to control group. Lin28 and Sall4 genes were significantly upregulated (P<0.0001, P<0.0001), whereas Dazl was upregulated (P<0.0001) after SSCs cryopreservation in alginate hydrogel.
Conclusion: This study indicates that use of alginate as scaffold in SSCs cryopreservation can be effective for stemness preservation of these cells.
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