title: Differentiation of Mouse Ovarian Stem Cells Toward Oocyte-Like Structure by Coculture with Granulosa Cells.
abstract: An increasing body of evidence has confirmed existence and function of ovarian stem cells (OSCs). In this study, a novel approach on differentiation of OSCs into oocyte-like cells (OLCs) has been addressed. Recently, different methods have been recruited to isolate and describe aspects of OSCs, but newer and more convenient strategies in isolation are still growing. Herein, a morphology-based method was used to isolate OSCs. Cell suspension of mouse neonatal ovaries was cultured and formed colonies were harvested mechanically and cultivated on mouse embryonic fibroblasts. For differentiation induction, colonies transferred on inactive granulosa cells. Results showed that cells in colonies were positive for alkaline phosphatase activity and reverse transcription-polymerase chain reaction (RT-PCR) confirmed the pluripotency characteristics of cells. Immunofluorescence revealed a positive signal for OCT4, DAZL, MVH, and SSEA1 in colonies as well. Results of RT-PCR and immunofluorescence confirmed that some OLCs were generated within the germ stem cell (GSCs) colonies. The applicability of morphological selection for isolation of GSCs was verified. This method is easier and more economic than other techniques. Our results demonstrate that granulosa cells were effective in inducing the differentiation of OSCs into OLCs through direct cell-to-cell contacts.
authors.affiliation: 1 Department of Anatomy, Faculty of Medicine, Alborz University of Medical Sciences , Karaj, Iran .
authors.forename: Soraya
authors.initials: S
authors.lastname: Parvari
authors.affiliation: 2 Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences , Tehran, Iran .
authors.forename: Hossein
authors.initials: H
authors.lastname: Yazdekhasti
authors.affiliation: 2 Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences , Tehran, Iran .
authors.forename: Zahra
authors.initials: Z
authors.lastname: Rajabi
authors.affiliation: 3 Department of Iranian Traditional Medicine, Tehran University of Medical Sciences , Tehran, Iran .
authors.forename: Valliollah
authors.initials: V
authors.lastname: Gerayeli Malek
authors.affiliation: 2 Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences , Tehran, Iran .
authors.forename: Tayebeh
authors.initials: T
authors.lastname: Rastegar
authors.affiliation: 2 Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences , Tehran, Iran .
authors.forename: Mehdi
authors.initials: M
authors.lastname: Abbasi
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques; D005260:Female; D006107:Granulosa Cells / Q000166:cytology*; D051379:Mice; D009865:Oocytes / Q000166:cytology*; D010053:Ovary / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*
keywords: coculture; infertility treatment; oocyte-like cells; ovarian stem cells