Tissue Culture To Observe Germ Cell Development In Immature Testis

Objective Established the tissue culture model of the immaturetestis, evaluating the effect of fetal bovine serum (FBS) and serumreplacement-KnockoutTMSR (KSR) on germ cell development andinvestigating the germ cell development in tissue culture of the humanimmature testicular tissue.Methodsâ‘ The FBS and KSR were used as supplement of culture medium, the SDtesticular tissues were cultured for5weeks. Area of the cultured tissueswas measured. The testicular histology, the development of the germcell and the diameter of seminiferous tubules were observed by HEstaining. The apoptotic cells were observed in the testis cultured for5weeks by apoptosis experiments. The various stages marker genes ofspermatogenesis, such as Kit, Sycp3and Crisp1, were identified byPT-PCR.â‘¡The testicular tissues of the testicular tumor para-carcunoma tissuewere used to tissue cultured. The testicular histology, the area and thediameter of seminiferous tubules were observed by HE staining after cultured4weeks-17weeks, respectively. The Piwil2gene was identifiedby PT-PCR, and the Piwil2protein expression was observed by theimmunohistochemistry.Resultsâ‘ In the KSR group, the cultured tissues cuold sustain growth and theseminiferous tubules gradually increased in the period of culture. Thespermatogonia, primary spermatocytes, secondary spermatocytes andround spermatids were observed in the cultured tissues after cultured5weeks, and their marker gene Kit, Sycp3and Crisp1were identified. Inthe FBS group, the testicular organization dwindled in the period ofculture; the seminiferous tubules appeared obviously necrosis and theexpression of the Sycp3and Crisp1were instability after cultured5weeks.â‘¡The seminiferous tubules area and the diameter significantly increasedin the early of the human tissue cultured (4weeks), and have adecreasing trend in the late period. The resembianc primaryspermatocytes were observed in the cultured tissues after cultured8weeks, Piwil2immunohistochemistry study suggests germ cells haveself-renewal ability.Conclusionsâ‘ KSR has more advantages for spermatogonial cells to develop intosperm cells in immature tissue culture, and can maintain the germ cell development and the cultured tissues growth development for a longtime.â‘¡The tissue culture system can maintain the germ cell development inthe humam immature testicular tissue. The germ cell still has the abilityof self-renewal and differentiation after cultured8weeks, and decliningof the germ cell in the cultured tissue.