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Roles for Osteocalcin in Proliferation and Differentiation of Spermatogonial Cells Cocultured With Somatic Cells Publisher Pubmed



Solhjoo S1 ; Akbari M1 ; Toolee H1 ; Mortezaee K2 ; Mohammadipour M3 ; Nematollahimahani SN4 ; Shahrokhi A5 ; Sayadi M6 ; Rastegar T1
Authors
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Authors Affiliations
  1. 1. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Department of Anatomy, School of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran
  3. 3. Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
  4. 4. Physiology Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran
  5. 5. Department of Pharmacy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
  6. 6. Department of Hematology, Allied Medical School, Tehran University of Medical Sciences, Tehran, Iran

Source: Journal of Cellular Biochemistry Published:2019


Abstract

Spermatogonial cells (SCs) are key cells for spermatogenesis. These cells are affected by paracrine signals originated from nearby somatic cells, among them Leydig cells have receptors for osteocalcin, a hormone known for exerting positive roles in the promotion of spermatogenesis. The aim of this study was to evaluate roles for osteocalcin on SCs proliferative and differentiation features after coculture with Leydig cells. SCs and Leydig cells were isolated from neonate NMRI offspring mice and adult NMRI mice, respectively. SCs population were then enriched in a differential attachment technique and assessed for morphological features and identity. Then, SCs were cocultured with Leydig cells and incubated with osteocalcin for 4 weeks. Evaluation of proliferation and differentiation-related factors were surveyed using immunocytochemistry (ICC), Western blot, and quantitative real-time polymerase chain reaction (PCR). Finally, the rate of testosterone release to the culture media was measured at the end of 4th week. Morphological and flow cytometry results showed that the SCs were the population of cells able to form colonies and to express ID4, α6-, and β1-integrin markers, respectively. Leydig cells were also able to express Gprc6α as a specific marker for the cells. Incubation of SCs/Leydig coculture with osteocalcin has resulted in an increase in the rate of expressions for differentiation-related markers. Levels of testosterone in the culture media of SCs/Leydig was positively influenced by osteocalcin. It could be concluded that osteocalcin acts as a positive inducer of SCs in coculture with Leydig cells probably through stimulation of testosterone release from Leydig cells and associated signaling. © 2018 Wiley Periodicals, Inc.
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