title: Short Term Culture of Vitrified Human Ovarian Cortical Tissue to Assess the Cryopreservation Outcome: Molecular and Morphological Analysis.
abstract: BACKGROUND
  The aim of the present study was to evaluate the effectiveness of human ovarian vitrification protocol followed within vitroculture at the morphological and molecular levels.
  
  
  METHODS
  Ovarian tissues were obtained from 10 normal transsexual women and cut into small pieces and were divided into non-vitrified and vitrified groups and some of the tissues fragments in both groups were randomly cultured for two weeks. The morphological study using hematoxylin and eosin and Masson's trichrome staining was done. The analysis of mean follicular density, 17-β estradiol (E2) and anti mullerian hormone (AMH), and real-time RT-PCR was down for the evaluation of expression of genes related to folliculogenesis. Data were compared by paired-samples and independent-samples T test. Values of p<0.05 were considered statistically significant.
  
  
  RESULTS
  The proportion of normal follicles did not show significant difference between vitrified and non-vitrified groups before and after culture but these rates and the mean follicle density significantly decreased in both cultured tissues (p<0.05). The expression of genes was similar in vitrified and non-vitrified groups but in cultured tissues the expression of GDF9 and FSHR genes increased and the expression of FIGLA and KIT-L genes decreased (p<0.05). An increase in E2 and AMH concentration was observed after 14 days of culture in both groups.
  
  
  CONCLUSIONS
  In conclusion, the present study indicated that the follicular development and gene expression in vitrified ovarian tissue was not altered before and afterin vitroculture, thus this method could be useful for fertility preservation; however, additional studies are needed to improve the culture condition.
authors.affiliation: Department of Anatomical Sciences, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Ramezani
authors.affiliation: Department of Anatomical Sciences, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran.
authors.forename: Mojdeh
authors.initials: M
authors.lastname: Salehnia
authors.affiliation: Reproductive Health Research Center, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Mina
authors.initials: M
authors.lastname: Jafarabadi
keywords: 17 beta- Estradiol; Anti- mullerian hormone; Gene expression; Vitrification