Basal lamina formation by porcine thyroid cells grown in collagen- and laminin-deficient medium.
Porcine thyroid cells isolated by dispase treatment were cultured in either (a) Matrigel, (b) agarose with the addition of different combinations of basic fibroblast growth factor and laminin, or (c) on agarose-coated dishes. The formation of follicles and the presence of a basal lamina was investigated by routine electron microscopy of Araldite-embedded material and by light and electron microscopical immunocytochemical detection of the basal lamina components, laminin and collagen type IV. After 10 days of culture in Matrigel or agarose, a basal lamina-like structure surrounded most follicles. Follicles of cells growing in agarose and overlaid with a medium containing thyrotropin and overlaid with a medium containing thyrotropin and fibroblast growth factor showed a fluorescent band at the basal side of the follicles after immunocytochemical staining with anti-laminin and anti-collagen antibodies. Routine electron microscopy showed that a basal lamina-like structure lined the outside of the follicle. This structure could be subdivided into a lamina lucida and a lamina densa. Electron microscopical immunogold labelling revealed that immunologically detectable laminin was confined to the lamina densa. These findings suggest that even in the absence of basal lamina components in the culture medium, thyroid cells are able to form follicles with a regular basal lamina when they are cultured in a three-dimensional environment.
Department of Anatomy, Eberhard-Karls University of Tübingen, Germany.
E
E
Fröhlich
R
R
Wahl
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K
Reutter
D000818:Animals; D001485:Basement Membrane / Q000378:metabolism; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000172:deficiency*; D003470:Culture Media; D004338:Drug Combinations; D005109:Extracellular Matrix / Q000378:metabolism; D007150:Immunohistochemistry; D007797:Laminin / Q000172:deficiency*; D008854:Microscopy, Electron; D011509:Proteoglycans; D012685:Sepharose; D013552:Swine; D013961:Thyroid Gland / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure
D003470:Culture Media; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D012685:Sepharose