title: Stem cell isolation by a morphology-based selection method in postnatal mouse ovary.
abstract: BACKGROUND
  An increasing body of evidence has emerged regarding the existence and function of spermatogonial stem cells (SSCs); however, their female counterparts are the subject of extensive debate. Theoretically, ovarian germ stem cells (GSCs) have to reside in the murine ovary to support and replenish the follicle pool during the reproductive life span. Recently, various methods have been recruited to isolate and describe aspects of ovarian GSCs, but newer and more convenient strategies in isolation are still growing. Herein, a morphology-based method was used to isolate GSCs.
  
  
  METHODS
  A cell suspension of mouse neonatal ovaries was cultured. Colonies of GSCs were harvested mechanically and cultivated on mouse embryonic fibroblasts (MEF). Alkaline phosphatase activity was assessed to verify stemness features of cells in colonies. Expression of germ and stem cell specific genes (Oct-4, Nanog, Fragilis, C-kit, Dazl, and Mvh) was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Immunofluorescence of Oct4, Dazl, Mvh, and SSEA-1 was also performed.
  
  
  RESULTS
  Small colonies without a clear border appeared during the first 4 days of culture, and the size of colonies increased rapidly. Cells in colonies were positive for alkaline phosphatase activity. Reverse transcription-polymerase chain reaction showed that Oct-4, Fragilis, C-kit, Nanog, Mvh, and Dazl were expressed in colony-forming cells. Immunofluorescence revealed a positive signal for Oct4, Dazl, Mvh, and SSEA-1 in colonies as well.
  
  
  CONCLUSIONS
  The applicability of morphological selection for isolation of GSCs was verified. This method is easier and more economical than other techniques. The availability of ovarian stem cells can motivate further studies in development of oocyte and cell-based therapies.
authors.affiliation: Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Soraya
authors.initials: S
authors.lastname: Parvari
authors.affiliation: Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Abbasi
authors.affiliation: Faculty of Medicine, Tehran Medical Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Niloufar
authors.initials: N
authors.lastname: Abbasi
authors.affiliation: Department of Iranian Traditional Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Valliollah Gerayeli
authors.initials: VG
authors.lastname: Malek
authors.affiliation: Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Fardin
authors.initials: F
authors.lastname: Amidi
authors.affiliation: Department of Anatomy, Faculty of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran.
authors.forename: Fereydoon Sargolzaei
authors.initials: FS
authors.lastname: Aval
authors.affiliation: Research Center, Iranian Blood Transfusion Organization (IBTO), Tehran, Iran.
authors.forename: Mehryar Habibi
authors.initials: MH
authors.lastname: Roudkenar
authors.affiliation: PrimeGen Biotech, Irvine, USA.
authors.forename: Fariburz
authors.initials: F
authors.lastname: Izadyar
keywords: isolation; mice; ovary; stem cells