Polarized epithelial cells of the hamster seminal vesicle in a serum-free bicameral culture system: evidence of secretory and endocytic activities.
Primary cultures of epithelial cells from the hamster seminal vesicle were established in a chemically defined medium supplemented with hormones and growth factors. Epithelial cell clusters were prepared combining enzymatic dissociation and mechanical disaggregation and then seeded in bicameral systems equipped with collagen-membrane inserts. A growth curve was generated and the cells were characterized morphologically and morphometrically by light and electron microscopy. The immunocytochemical detection of cytokeratins and the measurement of transepithelial electrical resistance were also performed. The secretory activity was studied by fluorography using L-[35S]methionine as a precursor, and endocytosis was approached using horseradish peroxidase as a tracer. Our results show that epithelial cells of the seminal vesicle can be grown as a monolayer of morphological and functionally polarized cells which retain secretory and endocytic activities. These cell cultures might therefore prove useful to investigate further the regulation of secretion and endocytosis in the seminal vesicle and are a promising model to approach, in a broader scope, cell polarity and protein sorting and targeting.
Rodrigues
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G
Department of Cell Biology, Gulbenkian Institute of Science, Apartado 14, 2781 Oeiras, Portugal.
Gulbenkian
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S
Mata
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LR
D000818:Animals; D002455:Cell Division / Q000502:physiology; D016764:Cell Polarity / Q000502:physiology*; D006224:Cricetinae; D016895:Culture Media, Serum-Free; D015194:Diffusion Chambers, Culture; D017097:Electric Impedance; D004705:Endocytosis / Q000502:physiology*; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D007150:Immunohistochemistry; D007633:Keratins / Q000032:analysis; D008297:Male; D008647:Mesocricetus; D012636:Secretory Rate; D012669:Seminal Vesicles / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure
D016895:Culture Media, Serum-Free; D007633:Keratins