In vitro effects of epidermal growth factor, follicle stimulating hormone and testosterone on mouse spermatogonial cell colony formation
S. Hadi Anjamrooz A , Mansoureh Movahedin A C , Taki Tiraihi A and S. Javad Mowla BA Department of Anatomical Sciences, School of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.
B Department of Genetics, School of Basic Sciences, Tarbiat Modarres University, Tehran, Iran.
C Corresponding author. Email: mansoure@modares.ac.ir
Reproduction, Fertility and Development 18(6) 709-720 https://doi.org/10.1071/RD05126
Submitted: 29 September 2005 Accepted: 9 April 2006 Published: 11 July 2006
Abstract
The complex process of spermatogenesis is regulated by various factors. In the present study, the in vitro effects of epidermal growth factor (EGF), follicle stimulating hormone (FSH) and testosterone on spermatogonial cell colony formation were investigated, and the best colonising factor was chosen for treating cells before transplantation. Sertoli and spermatogonial cells were isolated from neonatal mouse testes. The identity of the cells was confirmed through analysis of morphology, alkaline phosphatase activity, immunoreactivity and transplantation. Co-cultured Sertoli and spermatogonial cells were treated with EGF, FSH and testosterone before colony assay. Results indicated that EGF is the best factor for in vitro colonisation of spermatogonial cells, but transplantation of the EGF-treated group did not show any significant change compared with the control groups. In conclusion, EGF increased in vitro colonisation of spermatogonial cells, but, as a result of differential effects, did not influence transplantation efficiency.
Extra keywords: co-culture, colonisation, cytokine, spermatogenesis, spermatozoa.
Acknowledgments
This work was supported, in part, by a research grant from Tarbiat Modarres University.
Adekunle, A. O. , Falase, E. A. , Ausmanus, M. , Kopf, G. S. , Van-Arsdalen, K. N. , and Teuscher, C. (2000). Comparative analysis of blood plasma epidermal growth factor concentrations, hormonal profiles and semen parameters of fertile and infertile males. Afr. J. Med. Sci. 29, 123–126.
Bellve, A. R. , Cavicchia, J. C. , Millette, C. F. , O'Brien, D. A. , Bhatnagar, Y. M. , and Dym, M. (1977). Spermatogenic cells of the prepuberal mouse. Isolation and morphological characterization. J. Cell Biol. 74, 68–85.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Scarpino, S. , Morena, A. R. , Petersen, C. , Froysa, B. , Soder, O. , and Boitani, C. (1998). A rapid method of Sertoli cell isolation by DSA lectin, allowing mitotic analyses. Mol. Cell. Endocrinol. 146, 121–127.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Simoni, M. , Gromoll, J. , and Nieschlag, E. (1997). The follicle-stimulating hormone receptor: biochemistry, molecular biology, physiology, and pathophysiology. Endocr. Rev. 18, 739–773.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Singh, J. , and Handelsman, D. J. (1996). The effects of recombinant FSH on testosterone-induced spermatogenesis in gonadotrophin-deficient (hpg) mice. J. Androl. 17, 382–393.
| PubMed |
Spaliviero, J. A. , and Handelsman, D. J. (1991). Effect of epidermal and insulin-like growth factors on vectorial secretion of transferrin by rat Sertoli cells in vitro. Mol. Cell. Endocrinol. 81, 95–104.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Tajima, Y. , Sawada, K. , Morimoto, T. , and Nishimune, Y. (1994). Switching of mouse spermatogonial proliferation from the c-kit receptor-independent type to the receptor-dependent type during differentiation. J. Reprod. Fertil. 102, 117–122.
| PubMed |
Tfelt-Hansen, J. , Yano, S. , Macleod, R. J. , Smajilovic, S. , Chattopadhyay, N. , and Brown, E. M. (2005). High calcium activates the EGF receptor potentially through the calcium-sensing receptor in Leydig cancer cells. Growth Factors 23, 117–123.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Tsutsumi, O. , Kurachi, H. , and Oka, T. (1986). A physiological role of epidermal growth factor in male reproductive function. Science 233, 975–977.
| PubMed |
Uguralp, S. , Karabulut, A. B. , Mizrak, B. , Kaymaz, F. , Kiziltay, A. , and Hasirci, N. (2004). The effect of sustained and local administration of epidermal growth factor on improving bilateral testicular tissue after torsion. Urol. Res. 32, 323–331.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Wahab-Wahlgren, A. , Martinelle, N. , Holst, M. , Jahnukainen, K. , Parvinen, M. , and Soder, O. (2003). EGF stimulates rat spermatogonial DNA synthesis in seminiferous tubule segments in vitro. Mol. Cell. Endocrinol. 201, 39–46.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Wong, R. W. , Kwan, R. W. , Mak, P. H. , Mak, K. K. , Sham, M. H. , and Chan, S. Y. (2000). Overexpression of epidermal growth factor induced hypospermatogenesis in transgenic mice. J. Biol. Chem. 275, 18 297–18 301.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Yan, Y. C. , Sun, Y. P. , and Zhang, M. L. (1998). Testis epidermal growth factor and spermatogenesis. Arch. Androl. 40, 133–146.
| PubMed |
Yoshinaga, K. , Nishikawa, S. , Ogawa, M. , Hayashi, S. , Kunisada, T. , Fujimoto, T. , and Nishikawa, S. (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.
| PubMed |