اسپرماتوژنز فرایند پیچیدهای است که از سلولهای اسپرماتوگونی منشا گرفته و شامل مراحل پی در پی و سازمان یافتهای از تکثیر وتمایز سلولی است که در نهایت منجر به تولید سلولهای عملکردی اسپرماتوزوا می شود. جهت انجام این مطالعه ، نمونه گیری سلولهای سرتولی و اسپرماتوگونی از گوسالههای 5-3 ماهه انجام گرفت. در گروههای درمانی، کشت همزمان سلولهای سرتولی و اسپرماتوگونی، قبل از ارزیابی کلونی تحت درمان با هورمون گنادوتروپین کوریونی انسانی (hCG: human Chorionic Gonadotropin) 2، 5 و 10 واحد در هر میلی‌لیتر محیط کشت قرار گرفتند .برای شناسایی سلولهای سرتولی از رنگ‌آمیزی ایمونوسیتوشیمی ویمنتینن و کلونیهای اسپرماتوگونی بنیادی با روش ایمونوسیتوشیمی OCT-4 شناسایی شدند. نتایج نشان داد که شناسایی کلونیهای سلولهای بنیادی اسپرماتوگونی و سرتولی به خوبی به ترتیب توسط رنگ آمیزی ایمونوسیتوشیمی OCT-4 و ویمنتین انجام گرفته و هورمون hCG تعداد و قطر کلونی را افزایش می‌دهد این مطالعه نشان داد که .hCG می‌تواند منجر به القای تکثیر و فرایند تمایز در کشت سلولهای بنیادی اسپرماتوگونی و سرتولی شود.
Effects of human chorionic gonadotropin on spermatogonial stem cell colony formation and differentiation after in vitro Coculture with Sertoli cells
نویسندگان [English]
Narenji Sani, R., Tajik, P., Damghanaian, N., Javdani, G. .
.
چکیده [English]
Spermatogenesis is a complex developmental process that originates from spermatogonia stem cell. This process consists of sequential, highly organized steps of cell proliferation and differentiation resulting in the generation of functional spermatozoa. Therefore, the aim of the present study was to determine the effects of human chorionic gonadotropin (hCG) on spermatogonial stem cell colony formation and differentiation after in vitro Coculture with Sertoli cells. In this experimental study, Sertoli and spermatogonial cells were isolated from 3- to 5-month-old calves. Co-cultured Sertoli and spermatogonial cells were treated with hCG in treatment groups before colony assay with 2, 5 and 10 IU/ml culture medium. Vimentin and oct-4 immunohistochemical staining were used for Sertoli and spermatogonial stem cell colony detection, respectively. The present study showed that hCG increase both colony diameter and colony number. hCG can induce both proliferative pathway and differentiation process in SSCs Coculture with Sertoli cells.
Anway, M.D., Folmer, J., Wright, W.W., Zirkin, B.R. (2003): Isolation of Sertoli Cells from Adult Rat Testes: An Approach to Ex Vivo Studies of Sertoli Cell Function. Biol. Reprod. 68: 996-1002.
Baarends, W., Grootegoed, J., 1999. Molecular biology of male gametogenesis. Molecular biology in reproductive medicine. New York, USA: Parthenon Publishing Group, 271-295.
De Franca LR, L.R., Avelar, G.F., Almeida, F.F.L. (2005): Spermatogenesis and sperm transit through the epididymis in mammals with emphasis on pigs. Theriogenology 63: 300-318.
De Franca LR, L.R., Silva, V.A., Chiarini-Garcia, H.l., Garcia, S.K., Debeljuk, L. (2000): Cell Proliferation and Hormonal Changes During Postnatal Development of the Testis in the Pig. Biol. Reprod. 6: 1629-1636.
Dym, M., Cavicchia, J.C. (1977): Further Observations on the Blood-Testis Barrier in Monkeys. Biol. Reprod. 17: 390-403.
Enders, G.C., May Ii, J.J. (1994): Developmentally Regulated Expression of a Mouse Germ Cell Nuclear Antigen Examined from Embryonic Day 11 to Adult in Male and Female Mice. Develop. Biol. 163: 331-340.
Izadyar, F., Matthijs-Rijsenbilt, J.J., Ouden, K.D., Creemers, L.B., Woelders, H., de Rooij, D.G. (2002): Development of a Cryopreservation Protocol for Type A Spermatogonia. J. Androl. 23: 537-545.
Kojima, Y., Kominami, K., Dohmae, K., Nonomura, N., Miki, T., Okuyama, A., Nishimune, Y., Okabe, M. (1997). Cessation of Spermatogenesis in Juvenile Spermatogonia! Depletion (jsd/jsd) Mice. Inter. J. Urol. 4: 500-507.
Kubota, H., Avarbock, M.R., Brinster, R.L. (2004): Growth factors essential for self-renewal and expansion of mouse spermatogonial stem cells. Proceedings of the Nation. Acad. Sci. U. S. A. 101: 16489-16494.
Kumar, T.R (2005). What have we learned about gonadotropin function from gonadotropin subunit and receptor knockout mice? Reproduction. 130: 293-302.
Lamb, D.J., Spotts, G.S., Shubhada, S., Baker, K.R. (1991): Partial characterization of a unique mitogenic activity secreted by rat Sertoli cells. Mol. Cell. Endocrinol. 79: 1-12.
McLean, D.J., Russell, L.D., Griswold, M.D. (2002): Biological Activity and Enrichment of Spermatogonial Stem Cells in Vitamin A-Deficient and Hyperthermia-Exposed Testes from Mice Based on Colonization Following Germ Cell Transplantation. Biol. Reprod. 66: 1374-1379.
Meehan, T., Schlatt, S., O'Bryan, M.K., de Kretser, D.M., Loveland, K.L. (2000): Regulation of Germ Cell and Sertoli Cell Development by Activin, Follistatin, and FSH. Develop. Biol. 220: 225-237.
Pierce, J.G., Parsons, T.F. (1981): Glycoprotein hormones: structure and function. Ann. Rev. Bioch. 50: 465-495.
Scarpino, S., Morena, A.R., Petersen, C., Fröysa, B., Söder, O., Boitani, C. (1998): A rapid method of Sertoli cell isolation by DSA lectin, allowing mitotic analyses. Mol. Cell. Endocrinol. 146(1): 121-7.
Tajik, P., Sani, R.N., Moezifar, M., Yousefi, M., Movahedin, M., Qasemi-Panahi, B., Shafiei, S., Fili, P.R. (2014): Effect of follicle-stimulating hormone and testosterone on colony formation of bovine spermatogonial stem cell. Comp. Clin. Pathol. 23: 901-906.
17. Yoshinaga, K., Nishikawa, S., Ogawa, M., Hayashi, S., Kunisada, T., Fujimoto, T. (1991): Role of c-kit in mouse spermatogenesis: identification of spermatogonia as a specific site of c-kit expression and function. Development 113: 689-699.
ابتدای صفحه