Biopolymer gels as a basis of cryoprotective medium for testicular tissue of rats
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Biopolymer gels as a basis of cryoprotective medium for testicular tissue of rats Nataliia Volkova
. Mariia Yukhta . Anatoliy Goltsev
Received: 10 September 2018 / Accepted: 17 November 2018 Ó Springer Nature B.V. 2018
Abstract Cryopreservation of testis tissue is a promising approach to save fertility in prepubertal boys under going gonadotoxic cancer therapies. The using biopolymers as a basis of cryoprotective medium can be effective for the optimization of cryopreservation protocols of immature testicular tissue. The research purpose was to determine morphological parameters and metabolic activity of seminiferous tubules of immature rat testes under exposure to cryoprotective solution (DMSO) based on collagen or fibrin gels (CG or FG) as one of the first stages of developing the cryopreservation protocol. It was found that 30-min exposure of tissue samples to CG and FG with 0.6 M DMSO did not impair the spermatogenic epithelium and metabolic activity of the cells (MTT test and total lactate dehydrogenase activity). The use of FG at the time of exposure of 45 min did not lead to significant changes in the metabolic activity in contrast to other groups. The findings could be used to substantiate and develop the effective techniques for cryopreservation of immature seminiferous tubules.
N. Volkova (&) M. Yukhta A. Goltsev Department of Cryopathophysiology and Immunology, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv, Ukraine e-mail: [email protected]
Keywords Immature testicular tissue Spermatogenic epitelium Cryoprotective medium Collagen gel Fibrin gel Metabolic activity
Introduction In prepubertal boys affected by cancer, cryopreservation of testicular fragments prior to gonadotoxic treatment is an ethically accepted strategy to save fertility (Abrishami et al. 2010; Michele et al. 2017). Frozen immature testicular tissue can later be used for completion of spermatogenesis after in vitro maturation, germ cell transplantation or testicular tissue grafting (Travers et al. 2011). Despite the fact that the low temperature preservation of testis is associated with some difficulties (different cell size, post thawing ischemia), it can allow to preserve the different testicular cells in their ‘‘niche’’ with respect interactions between germ cells and Sertoli cells (Milazzo et al. 2010). Nevertheless, the optimization of cryopreservation protocols for improving recovery has not been practically investigated. In our opinion, the various biotechnological methods using natural biopolymers can be effective for this purpose, but they require additional studies. Tissue fragment encapsulation is based on cell scaffold technology (Nicodemus and Bryant 2008). The latter relies on immobilization of cells in a
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Cell Tissue Bank
biomaterial which allows bidirectional diffusion of nutrients, oxygen, and waste, thus promoting cell interactions. Substances for encapsulation should also be bioco
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