Knock-Out Serum Replacement and Melatonin Effects on Germ Cell Differentiation in Murine Testicular Explant Cultures

Ahmed Reda, Halima Albalushi, Sheyla Cisneros Montalvo, Mirja Nurmio, Zeliha Sahin, Mi Hou, Niels Geijsen, Jorma Toppari, Olle Söder, Jan Bernd Stukenborg*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

24 Citations (Scopus)


Finding robust culture conditions for in vitro maturation (IVM) of male germ cells is still a challenge. Recently, a testis organ culture method, using Knockout Serum Replacement (KSR), was suggested as a promising approach. However, the efficiency of that model is still not optimal. Hence, we have tried to establish the culture conditions in two laboratories, and to improve the reliability of the culture system to generate mature germ cells. Male mice at three days of age were sacrificed. Testes were cut into small pieces which were cultured atop agarose stands, using Minimum Essential Medium alpha supplemented with different supplements; melatonin, Glutamax, and different concentrations of KSR. The results showed that the duration of culture beyond 18 days had an impact on the number of differentiated germ cells. Supplementation with melatonin and Glutamax revealed a positive influence on the efficiency of male germ cell differentiation in vitro. Furthermore, the results confirmed that KSR had a positive effect on germ cell maturation and testosterone production, with a concentration of at least 10%. In conclusion, this study emphasizes the beneficial role of at least 10% KSR in the IVM of germ cells.

Original languageEnglish
Pages (from-to)1783-1794
Number of pages12
JournalAnnals of Biomedical Engineering
Issue number7
Publication statusPublished - Jul 1 2017


  • CREM
  • DDX4
  • Germ cell
  • In vitro spermatogenesis
  • KSR

ASJC Scopus subject areas

  • Biomedical Engineering


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