Different associations of cryoprotectants for testicular tissue of prepubertal cats submitted to vitrification

Abstract

The cryopreservation of testicular tissue is presented as the only alternative for the preservation of genetic material from prepubertal animals. However, this biotechnology is still being tested. The objective of this study was to evaluate the effect of different associations of cryoprotectants and the potential of cell proliferation after vitrification of testicular tissue of prepubertal cats. Five testicular pairs from five prepubertal cats were used, and each pair was divided into four fragments. Of these, one fragment composed of the control group (CG) and the rest were distributed in experimental groups according to the associations of cryoprotectants to be tested (dimethyl sulphoxide (DMSO)/glycerol (GLY); ethylene glycol (EG)/GLY) or DMSO/EG) in a final cryoprotectant concentration of 5.6 m. The fragments were submitted to vitrification, and after one week, fragments were heated and processed for histomorphological evaluation and quantification of nucleolar organizer regions (NORs). DMSO/GLY did not differ from CG and was superior to the other vitrified groups, as to cell separation and degree of shrinkage of the basal membrane. Concerning cell differentiation, visibility of the nucleus and nuclear condensation, all the vitrified groups were inferior to CG; however, DMSO/EG was inferior to DMSO/GLY and EG/GLY, which did not differ among themselves. CG was superior to all groups in quantification of NORs. DMSO/EG was inferior to all others, and there was no difference between DMSO/GLY and EG/GLY. The association DMSO/GLY presented the best preservation of tissue integrity and potential of cell proliferation after vitrification of the testicular tissue of prepubertal cats.

Influence of warming and reanimation conditions on seminiferous tubule morphology, mitochondrial activity, and cell composition of vitrified testicular tissues in the domestic cat model

“…Long-term preservation of testicular tissues provides more options to maintain genetic diversity and sustainability in populations of rare and endangered species. Despite other techniques available for pubertal animals (including the rescue of epididymal sperm cells) to subsequently transmit genes to the next generation, testicular tissue is the only biomaterial for preserving the fertility of prepubertal animals that died unexpectedly [ 1 , 2 ]. In addition, oncological treatments for young boys are gonadotoxic and may lead to infertility [ 3 ].…”

“…Thus, the cryopreservation of testicular tissue can be an option to preserve the fertility of these cancer patients [ 4 ]. Previous studies aiming at establishing optimal protocols to protect biological material for future utilization in assisted reproduction have been conducted in different species such as mouse [ 5 ], humans [ 6 ], cats [ 1 ], dogs [ 7 ], and wild species [ 8 ]. However, a lot of progress remains to be done in male fertility preservation.…”

“…Warming samples at 37°C for 3 minutes or 97–100°C for 30–40 seconds led in both cases to >85% of cell viability. Warming at 37°C for 1 minute have been used for vitrified testicular tissue in different species, including cats [ 1 , 10 ]. Although quick exposure to high temperatures is beneficial for oocytes [ 16 ], warming conditions using similar approaches have never been evaluated in testicular tissues, including in domestic cats.…”

Cryopreserved biopsy tissues of rectal cancer liver metastasis for assessment of anticancer drug response in�vitro and in�vivo

“…Compared with the PDX model, tissue slice cultivation represented a more convenient and practical model in light of the shorter culture cycle and simpler operation. Certainly, the insert support was helpful for maintaining slice viability, and enabled the implementation of a steady platform and provided an ample supply of oxygen and nutrition, compared with floating alone (18,26,27). On the whole, precision-cut slice cultivation retained the heterogeneous architecture of the original tumor and thus might provide a basis for in-depth study of pharmacokinetic mechanisms in pharmacology and gene pathways in tumor biology.…”

Cryopreservation of testicular tissue: an alternative to maintain the reproductive capacity in different animal species

“…After the testicular tissue is exposed to the vitrification solution, fragments are placed in a metal cube held above liquid nitrogen in an insulated box, which enables the tissue to be cooled ultrarapidly. Upon cooling, these fragments are placed in cryotubes and kept in liquid nitrogen (LIMA et al, 2016). Among all the testicular cryopreservation techniques, vitrification is the one with the lowest operating costs and is the easiest to carry out.…”