Production of fibronectin and collagen types I and III by chick embryo dermal cells cultured on extracellular matrix substrates.
Dermal cells isolated from the back skin of 7-day chick embryos were cultured on homogeneous two-dimensional substrates consisting of one or two extracellular matrix components (type I, III, or IV collagen, fibronectin and several glycosaminoglycans (GAGs): hyaluronate, chondroitin-4, chondroitin-6, dermatan and heparan sulfates). The effect of these substrates on the production of fibronectin, of types I, III and IV collagen by cells was compared with that of culture dish polystyrene. Using immunofluorescent labeling of cultured cells, it was observed that, on all substrates, in 1-day and 7-day cultures, 85 to 95% of cells contain type I collagen in the perinuclear cytoplasm; label was absent from cell processes. Type I collagen was also detected in extracellular fibers extending between neighboring cells. By contrast, on all substrates, only 5 to 20% of cells produced type III collagen. Otherwise distribution of type III collagen was similar to that of type I collagen. With anti-type IV collagen antibody no staining of either cell content or extracellular spaces was detected. Staining with anti-fibronectin antibody revealed two types of distribution patterns. On polystyrene and on all but type I collagen substrates, labeling revealed clusters of short thick strands and patches of fibronectin-rich material in extracellular spaces. On type I collagen substrate, however, immunostaining revealed a delicate network of regularly spaced parallel fibrils of fibronectin extending between and along cells. Using quantitative radioimmunoassay of the culture media, it was shown that, after 7 days of culture, cells secreted more type I than type III collagen.(ABSTRACT TRUNCATED AT 250 WORDS)
Robert
J
J
Département de Biologie, Université Joseph Fourier, Grenoble, France.
Hartmann
D J
DJ
Sengel
P
P
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D003094:Collagen / Q000096:biosynthesis* / Q000378:metabolism; D005109:Extracellular Matrix / Q000502:physiology*; D005353:Fibronectins / Q000096:biosynthesis*; D005455:Fluorescent Antibody Technique; D011863:Radioimmunoassay; D012867:Skin / Q000166:cytology / Q000196:embryology* / Q000378:metabolism
D005353:Fibronectins; D003094:Collagen