Two- and three-dimensional system in agarose matrix for in vitro culture of testicular fragments of adult sheep

The restoration of spermatogenesis through approaches such as in vitro culture (IVC) of testicular tissue has been investigated as an alternative for the resumption of male fertility. However, the IVC system of testis from adult animals is still not well established. Therefore, the present study aimed to compare the two- (2D) and three-dimensional (3D) systems in an agarose matrix for the IVC of testicular fragments from adult sheep. For this, tissue histomorphology, cell proliferation, as well as fibrosis formation after 48 h of IVC were evaluated. Morphological analysis between the two systems revealed that despite cell loss and basement membrane rupture in the 2D system were higher, the 3D system showed more nuclear condensation. In addition, the 3D system showed less potential for cell proliferation and signs of tissue fibrosis. In conclusion, although, both IVC systems (2D and 3D) for testicular fragments showed morphological changes in the seminiferous tubules, the 2D system was better, as cell proliferation and collagen fibers were similar to fresh fragments.