Short Term Culture of Vitrified Human Ovarian Cortical Tissue to Assess The Cryopreservation Outcome: Molecularand Morphological Analysis

Background: The aim of the present study was to evaluate theeffectiveness of human ovarian vitrification protocol followedwith in vitro culture at the morphological and molecular levels.Materials and Methods: Ovarian tissues were obtained from۱۰ normal transsexual women and cut into small pieces andwere divided into non-vitrified and vitrified groups and someof the tissues fragments in both groups were randomly culturedfor two weeks. The morphological study using hematoxylin andeosin and Masson's trichrome staining was done. The analysisof mean follicular density, ۱۷-β estradiol (E۲) and anti mullerianhormone (AMH) and real-time RT-PCR was down forthe evaluation of expression of genes related to folliculogenesis.Data were compared by paired-samples and independentsamplest test. Values of P<۰.۰۵ were considered statisticallysignificant.Results: The proportion of normal follicles did not show significantdifference between vitrified and non-vitrified groups beforeand after culture but these rates and the mean follicle densitysignificantly decreased in both cultured tissues (P<۰.۰۵).The expression of genes was similar in vitrified and non-vitrifiedgroups but in cultured tissues the expression of GDF۹ andFSHR genes increased and the expression of FIGLA and KIT-Lgenes decreased (P<۰.۰۵). An increase in E۲ and AMH concen tration was observed after ۱۴ days of culture in both groups.Conclusion: The present study indicated that the follicular developmentand gene expression in vitrified ovarian tissue wasnot altered before and after in vitro culture, thus this methodcould be useful for fertility preservation; however, additionalstudies are needed to improve the culture condition.

کلیدواژه ها:

۱۷ Beta- Estradiol ، Anti-Mullerian Hormone ، Gene Expression ، Vitrification