ObjectiveTo evaluate the efficiency of two vitrification protocols for rat immature
testicular tissue and heterotopic transplantation.MethodsTwenty-four pre-pubertal Wistar rats were divided into three groups (n=8).
After orchiectomy, testicular fragments (3mm) from Groups 1 and 2 were
vitrified with different cryoprotectant concentration solutions, using
sterile inoculation loops as support. After warming up, the fragments were
submitted to cell viability assessment by Trypan blue and histological
evaluation. Vitrified (Groups 1 and 2) and fresh (Group 3) fragments were
grafted to the animals periauricular region. After 8 weeks of grafting, the
implant site was histologically analyzed.ResultsThe viability recovery rate from Group 1 (72.09%) was higher
(p=0.02) than that from Group 2 (59.19%). Histological
analysis showed similar tubular integrity between fresh fragments from
Groups 1 and 3. Group 2 samples presented lower tubular integrity. We ran
histological analyses in the grafts from the Groups. In all groups, it was
possible to see the implant site, however, no fragment of testicular tissue
or signs of inflammation were histologically found in most samples from
Groups 1 and 3. In one sample from Group 2, we found degenerated
seminiferous tubules with necrosis and signs of an inflammatory process. In
another sample from Group 2, we found seminiferous tubules in the implant
site.ConclusionThe vitrification of pre-pubertal testicular tissue of rats showed little
damage to cell viability through histological analysis when we used
cryoprotectants in a lower concentration. Heterotopic transplantation could
not preserve the structural organization of the testicular tissue.
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