The effects of fetal serum and gonadotropins on testosterone production by cultured chick testicular cells

• Published in: Domestic animal endocrinology Vol. 11; no. 3; pp. 261 - 269
• Main Authors: Castro, J.I., Romano, M.C.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.1994
• Subjects: Animals; Cells, Cultured; Chickens; Chorionic Gonadotropin - pharmacology; Culture Media; Fetal Blood; Follicle Stimulating Hormone - pharmacology; Gonadotropins - pharmacology; Luteinizing Hormone - pharmacology; Male; Testis - drug effects; Testis - metabolism; Testosterone - biosynthesis
• ISSN: 0739-7240; 1879-0054
• DOI: 10.1016/0739-7240(94)90017-5

This report describes the influences of fetal serum and gonadotropins on the development and function of chick testicular cells in a culture system that allows long term control of culture conditions. Testis of new born male chicks were dissociated and cultured in different conditions and the production of testosterone measured by RIA. 1. 1.|To investigate the effect of serum the cells were wither cultured in a defined medium composed by Dulbecco's modified medium (DMM) plus 0.1% albumin or media containing DMM plus 10% fetal bovine serum (FBS). The presence of serum increased protein content of the cultures, but no differences were found in basal testosterone production. When hCG (2IU/ml) was added to both culture media, the increment in testosterone production hence produced, was greater when cells were cultured in defined medium. 2. 2.|Cells were precultured for 24 hr in either defined serum media with or without hCG. The media were then replaced with DMM plus BSA and 1-methyl 3-isobutyl xanthine (IBMX) in the presence or absence of hCG and cells further incubated for 2 hr. Addition of hCG increased testosterone production by cells cultured in defined medium and enhanced its subsequent response to hCG stimulation. Cells cultured in serum medium maintained their ability to respond to hCG, but the addition of this hormone to the medium, abolished the response to further hCG-stimulation. 3. 3.|hCG caused a dose-related increased in testosterone production by cells cultured in defined and serum containing media, but concentrations of hCG above 2 IU/ml depressed testosterone production in the latter group. 4. 4.|Cells cultured in defined medium were incubated for 6 hr either with or without FSH of different purity grade. Both preparations stimulated the production of testosterone, the pure one being less effective than the LH-contaminated one. 5. 5.|Cells cultured in defined medium were incubated for 6 hr either with or without FSH and further stimulated with hCG (2 IU/ml). A significant increment in the response to hCG was observed when cells had been exposed to FSH. Present results show that FBS influences the function and development of chick testis cells in culture. Since both FSH and hCG modified testosterone secretion, a role for gonadotropins in the functional capacity of neonatal Leydig cells is also suggested.