Morphological aspects of cultured rat seminiferous tubules in the presence of fetal calf serum and follicle-stimulating hormone.
Culturing seminiferous tubules allows the analysis of spermatogenesis under controlled conditions. Reproducing the specific microenvironment for germ cells is a challenge taken up by various studies. The difficulty in supplementing all nutrients and the physical disturbances during isolation procedures cause degeneration of many cells in the seminiferous epithelium. We tested some culture conditions in order to preserve an acceptable morphology of cells inside the tubules. Seminiferous tubules were cultured during fifteen days in HAM F10 medium supplemented with fetal calf serum (FCS) and/or follicle-stimulating hormone (FSH). The cellular morphology was analyzed using scanning and transmission electron microscopy. During the culture period cellular degeneration occurred progressively. Morphological modifications of the Sertoli cells and germ cells such as accumulation of lipid droplets, nuclear and cytoplasmatic vacuolization and the presence of cell debris were observed. The addition of FCS activated the myoid cells causing nuclear rounding and thickening of the tubular wall. The best results were obtained with a serum-free culture medium supplemented with FSH.
Lombello
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CB
Department of Cellular Biology, Biology Institute, State University of Campinas, UNICAMP, São Paulo, Brazil. clombello@hotmail.com
Wada
M L
ML
Dolder
H
H
D000818:Animals; D001769:Blood; D003470:Culture Media; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D008297:Male; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D009924:Organ Culture Techniques; D051381:Rats; D017208:Rats, Wistar; D012671:Seminiferous Tubules / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure*
D003470:Culture Media; D005640:Follicle Stimulating Hormone