Study of the gynecological organs and breasts of rats submitted to cryopreserved ovarian transplantation and treated with adipose tissue derived stem cells

Abstract

The avascular ovarian transplantation is a promising approach for female fertility preservation, however the major concern is the initial hypoxia, until the reestablishment of a new vascular network. The adipose-tissue derived stem cells (ASCs) therapy has been proposed to enhanced angiogenesis in hypoxic tissues. This study aims to assess gynecological organs and breasts of rats submitted to cryopreserved ovarian transplantation and treated with ASCs. Twenty adults female Wistar rats will be submitted to bilateral oophorectomy. Both ovaries will be cryopreserved by slow freezing protocol and maintained in N2 storage for 24h. After this period, the ovaries will be thawed and a bilateral autologous transplantation with the whole ovaries without vascular anastomosis in the retroperitoneum will be performed. At this time, the animals will be divided into two groups: Gelfoam (Control) and Gelfoam + ASCs (5x104 cells/ovary). In Control group, the Gelfoam will be soaked in culture medium. Euthanasia will be held between 30-35 postoperative day, always in diestrus phase of the estrous cycle. The first pair of inguinal mammary glands, uterus and vagina will be removed for the following analysis: histo-morphological, morphometric and immunohistochemical for cell proliferation (Ki-67) and hormone receptors of progesterone and estrogen.