title: Mouse preantral follicle development in two-dimensional and three-dimensional culture systems after ovarian tissue vitrification.
abstract: OBJECTIVE
  This work on follicle culture and evaluation of expression of oocyte maturation genes helps to better understand the complicated processes of folliculogenesis and to develop new approaches for infertility treatment.
  
  
  METHODS
  Ovaries of 12-day-old female NMRI mice were divided into control and vitrification groups. After vitrification and warming procedures, ovarian tissue morphologies were histologically evaluated and compared to those of the control group. In the second stage, preantral follicles were mechanically isolated from non-vitrified and vitrified ovaries and cultured for 12 days in two-dimensional (2D) and three-dimensional (3D) systems. Finally, the survival and growth rate of follicles and quantitative expression of oocyte maturation genes (Gdf9, Bmp15 and Bmp6) were studied.
  
  
  RESULTS
  Morphological integrity of ovarian tissue in vitrification group was well preserved. Survival rates of cultured preantral follicles in control group during 2D and 3D systems were somewhat similar, but were significantly different between 2D and 3D systems in vitrification group. Although the growth rate of follicles was similar in the 3D system in both groups, substantially higher growth rate was observed for the control group in the 2D system. Expressions of oocyte maturation genes were, to some extent, similar between control and vitrification groups. There was a remarkable reduction in expression pattern of genes in 3D compared to 2D system in both experimental groups, during the 12th day of culture period.
  
  
  CONCLUSIONS
  3D in-vitro culture system could be appeared more appropriate than 2D culture system for preservation of follicles in terms of spatial morphology, growth rate and expression reduction of maturation genes.
authors.affiliation: Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran; Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
authors.forename: Seyedeh Zeynab
authors.initials: SZ
authors.lastname: Sadr
authors.affiliation: Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran. Electronic address: b.ebrahimi@royaninstitute.org.
authors.forename: Bita
authors.initials: B
authors.lastname: Ebrahimi
authors.affiliation: Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Shahhoseini
authors.affiliation: Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
authors.forename: Roya
authors.initials: R
authors.lastname: Fatehi
authors.affiliation: Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
authors.forename: Raha
authors.initials: R
authors.lastname: Favaedi
mesh_terms: D000818:Animals; D055430:Bone Morphogenetic Protein 15 / Q000235:genetics; D055418:Bone Morphogenetic Protein 6 / Q000235:genetics; D005260:Female; D015870:Gene Expression; D055429:Growth Differentiation Factor 9 / Q000235:genetics; D051379:Mice; D009865:Oocytes / Q000502:physiology; D006080:Ovarian Follicle / Q000502:physiology*; D010053:Ovary / Q000166:cytology; D046509:Tissue Culture Techniques; D058989:Vitrification
chemical_list: C525902:Bmp15 protein, mouse; C525817:Bmp6 protein, mouse; D055430:Bone Morphogenetic Protein 15; D055418:Bone Morphogenetic Protein 6; C525898:Gdf9 protein, mouse; D055429:Growth Differentiation Factor 9
keywords: Oocyte maturation genes; Ovarian tissue; Preantral follicles; Three-dimensional system; Vitrification