Cellular and gene therapy in experimental fresh and cryopreserved ovary transplant

Abstract

Cryopreserved ovary transplant has proved to be effective in the preservation of fertility in young patients who are submitted to cytotoxic treatments. Nevertheless, follicular depletion, after the transplant, limits future reproduction since the time lag spent in obtaining adequate perfusion of the transplanted tissue is an important factor of follicular survival and the functional longevity of the graft. Considering the imminent clinical applicability of the ovarian transplant in order to preserve female fertility and the necessity for techniques that enhance and optimize its use, this work aims at studying the effect of two kinds of treatments in fresh and cryopreserved ovary transplant in female rats - cellular therapy with pre-adipocytes and gene therapy with pre-adipocytes transfected with vascular endothelial growth factor - VEGF. Adult female rats were distributed in four experimental groups plus six control groups: First step: treatment with pre-adipocytes, and Second step: treatment with pre-adipocytes transfected with VEGF. Preadipocytes will be obtained from humane adipose tissue. Serum dosages of stradiol and progesterone will be performed before and after the transplant, besides daily vaginal smears until after thirty days after the transplant when euthanasia will occur. Morphological and histomorphometric analyses will be performed as well as immunohistochemical analysis of cellular proliferation (Ki-67), apoptosis (caspasis-3) and neoangiogenesis (VEGF -A). PCR study of neoangiogenesis, apoptosis inflammation and oxidative stress will also be performed.