P-420 Involvement of Interluekin-6 (IL-6) in the development of different stages of spermatogenesis in vitro, using spermatogonial cells isolated from normal and busulfan-treated immature mice.

Abstract Study question Can Interluekin-6 (IL-6) induce spermatogonial cells from seminiferous tubules of normal or busulfan-treated mice to develop spermatogenesis in vitro using methylcellulose culture system (MCS)? Summary answer IL-6 induced the development premeiotic, meiotic and post-meiotic vitro. What is known already Spermatogenesis a complicated process sperm generation. During this proliferate differentiate meiotic, postmeiotic stages that continue spermiogenesis generate mature sperm. This under regulation autocrine paracrine factors provided by developed germ somatic such as Sertoli, peritubular Leydig cells. These microenvironmental are modified following pathological condition, chemotherapy, which may lead subfertility sterility. Different systems were used complete vitro; however, was not yet achieved. design, size, duration Sexually immature (7-day-old) intraperitoneally (i.p) injected with busulfan (45 mg/kg) isolate their (STs). Cells enzymatically isolated STs cultured (MCS), 3-dimensinal system. Fresh media without (CT) added cultures beginning, after two weeks. The determined 4 weeks Participants/materials, setting, methods Isolated (2x105/well/0.5ml) MCS contained StemPro-34 medium, KSR, rEGF, rGDNF, rLIF, r-bFGF presence/absence (1,10 100 pg/ml) incubated for four CO2 incubator at 37°C. colonies/organoids examined microscopically quantified different Sertoli functional markers immunofluorescence staining (IF) and/or qPCR analyses. Main results role chance Addition significantly increased premeiotic (VASA), (BOULE) meiotic/postmeiotic (ACROSIN) specific IF also differently affected expression levels cell functionality (androgen receptor, androgen binding protein, transferrin, GDNF, FSHR). Furthermore, addition VASA-, BOULE- ACROSIN-positive (as analyses). However, effect more potent compared mice. Also, distinctly Limitations, reasons caution in-vitro animal models. bio-activity needs be confirmed Wider implications findings indicate possible involvement could directly on through Our result suggest potential factor developing future therapeutic strategies male fertility preservation Trial registration number applicable