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.

Encapsulation of Satureja khuzistanica extract in alginate hydrogel accelerate wound healing in adult male rats

“…Alginate hydrogels are a kind of hydrogels based on natural polymers that are similar to extracellular matrices (ECM) so that they can participate in the wound healing process by using drug delivery. Therefore, alginate gels can play roles in functional improvement and release of bioactive compounds of drugs or plant materials in the wound site [14–17].…”

Testicular tissue cryopreservation is the preferred method to preserve spermatogonial stem cells prior to transplantation

“…Interestingly, longer colonies were observed after cryopreservation of TET compared with fresh controls or TCS. It has been reported that after exposing SSCs to freezing-thawing, stemness genes are up-regulated (Pirnia et al, 2016). Hence, during the initial period of colony expansion following SSCT, when spermatogonia proliferate circumferentially as well as along the tubule before engaging in spermatogenic differentiation (Nagano et al, 1999), frozen-thawed SSCs from TET may have propagated and colonized more tubule surface compared with fresh SSCs or SSCs from frozen-thawed TCS.…”

Advanced bioengineering of male germ stem cells to preserve fertility

“…Attractive features of this system include the ease of de-gelling process and retrieval of cells, transparency enabling for optical and fluorescence examination, their desirable porous network, and their limited inherent cell adhesion making them of interest for cell encapsulation and cell cultivation applications. 81 , 82 , 97 – 101 Alginate hydrogel properties such as desirable nutrient release, oxygen diffusivity, hydrophilicity, and antioxidant activity give them the potential to enhance cell survival and support their further proliferation. Jalayeri et al investigated the biocompatibility of alginate, encapsulating mouse spermatogonial stem cells.…”