Presentation Authors: Keykavos Gholami, Gholamreza Pourmand*, Morteza Koruji, Mohammadali Sadighigilani, Shadan Navid, Mehdi Abbasi, Tehran, Islamic Republic of Iran, Fariborz Izadyar, california, CA
Introduction: 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.
Methods: 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 theresults were compared with that of a two-dimensional (2D) culture system.
Results: We found that the number and diameter of colonies and the levels of expression of Scp3 and Integrin Î±6 in the 3D culture group were significantly higher than in the 2D group.
Conclusions: Our findings indicate that SACS can reconstruct a microenvironment capable of regulating both proliferation and differentiation of cell colonies.