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Efficiency of Colony Formation and Differentiation of Human Spermatogenic Cells in Two Different Culture Systems Publisher Pubmed



Gholami K1 ; Pourmand G2 ; Koruji M3 ; Sadighigilani M4 ; Navid S1 ; Izadyar F5 ; Abbasi M1
Authors
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Authors Affiliations
  1. 1. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Urology Research Center, Sina Hospital, Tehran University of Medical Sciences, Tehran, Iran
  3. 3. Cellular and Molecular Research Center & Department of Anatomical Sciences, Iran University of Medical Sciences, Tehran, Iran
  4. 4. Department of Urology, School Medicine, Tehran University of Medical Sciences, Tehran, Iran
  5. 5. PrimeGen Biotech, Irvine, CA, United States

Source: Reproductive Biology Published:2018


Abstract

Optimization of in vitro culture system for the expansion and the maturation of male germ cells to post meiotic stages is a valuable tool for studies exploring spermatogenesis regulation and the management of male infertility. Several studies have reported promising results of mouse spermatogonial stem cells culture in three-dimensional (3D) culture systems and a subsequent production of sperm. In the present study, we investigated the capacity of a three-dimensional soft agar culture system (SACS) supplemented with Knockout Serum Replacement (KSR) in colony formation and inducing human germ cells to reach post-meiotic stages. Testicular cells from testes of brain -dead donors were first cultured for three weeks in proliferation medium. The cells were subsequently cultured in the upper layer of the SACS (3D group) in a medium supplemented with KSR and hormones, and the results were compared with that of a two-dimensional (2D) culture system. We found that the number and diameter of colonies and the levels of expression of Scp3 and Integrin α6 in the 3D culture group were significantly higher than in the 2D group. Our findings indicate that SACS can reconstruct a microenvironment capable of regulating both proliferation and differentiation of cell colonies. © 2018 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn
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