ABSTRACT. The present study was aimed to examine the effects of follicle-stimulating hormone (FSH) on cell division of Sertoli cells from rat fetal testes and on the kinetics of 2 kinds of intermediate filaments, cytokeratin and vimentin, which comprise the cytoskeleton of Sertoli cells. Testes from rat fetuses of different ages (from day 15 to day 17 of gestation ) were cultured for 48 hr, with or without added FSH. for the presence of sperm in the vaginal smear. The day on which sperm was detected was counted as day 0 of gestation. Fetuses of 15-17 days of age were used for culture of their testes. These fetuses were removed from maternal uteri under aseptic conditions and transferred to sterile petri dishes containing Hanks' balanced salt solution (HBSS) with penicillin (100 units/ml) and streptomycin (100 µg/ml). Their testes were removed and rinsed with HBSS. Each testis was placed on the surface of a Millipore filter in the medium (Medium RPMI 1640; Gibco-BRL, U.S.A.) containing 0.2% sodium bicarbonate. Three series of culture were prepared. One was added with no FSH as control (group C). The second was added with 0.5 µg/ml FSH (Sigma, 1 mg equivalent to 1 unit shown by Steelman and Pohley [18]) as group FSH 0.5. The third was added with 5 µg/ml FSH as group FSH 5. The Millipore filter rested on a stainless-steel grid in a culture dish (Nunc, Roskilde, Denmark). All the cultures were incubated at 37˚C in a humidified atmosphere of 5% CO 2 and 95% O 2 for 48 hr. All of the above procedures were performed under sterile conditions.After incubation, the cultures were immediately frozen in liquid nitrogen. Frozen sections (8 µm) were air-dried for a few min, fixed in acetone at 20˚C for 10 min and allowed to be air-dried. The indirect immunofluorescence with amplification by the biotin-streptavidin system was used. Sections were incubated with monoclonal antibody to cytokeratin (clone Lu5, Boeringer Mannheim GmbH, Germany) or vimentin (clone V 9, Biodesign International, U.S.A.) and visualized with rhodamine X isothiocyanate (XRITC) or fluorescein isothiocyanate (FITC), respectively.To measure the proliferative activity of Sertoli cells in cultured testes, in another series of experiments, 10 µM 5-bromo-2'-deoxyuridine (BrdU) was added to the culture medium after 24 hr-incubation and the incubation was continued for 2 hr. Then, each explant was transferred Many studies have been made on differentiation and development of rat testes during the fetal period. In the formation of seminiferous tubule in the rat [20] and mouse [3], several mesenchyme-derived cells initially surround primordial germ cells to create a tube-like shape in the gonadal primordium. These mesenchyme-derived cells are considered to differentiate into Sertoli cells thereafter [7,8].It has been reported that intermediate filaments, cytokeratin and vimentin, are simultaneously found in Sertoli cells during the fetal period [15]. Cytokeratin disappears on 14 days after birth. Rat Sertoli cells show active cell division and proliferation during t...