Regulation of collagenase and collagenase mRNA production in early- and late-passage human diploid fibroblasts.
The levels of collagenase and collagenase mRNA produced by early-passage (less than 40% of lifespan completed) and late-passage (greater than 80% of lifespan completed) cultures of human fibroblasts were analyzed. The constitutive levels of collagenase and collagenase mRNA produced by the late-passage cultures were 10-30 x greater than the levels observed in similarly treated early-passage cultures. Immunofluorescence analysis established that the percentage of collagenase-positive cells was also greater (77% vs. 4%) in the late-passage cultures. To determine whether the difference in collagenase production resulted from cell-derived regulatory factors, collagenase production was examined in cultures plated onto substrates coated with fibroblast extracellular matrix (ECM). Collagenase and collagenase mRNA production was enhanced in both types of cultures, although amounts produced by ECM-induced early-passage cultures was significantly less than that produced by similarly treated late-passage cultures. Collagen-coated substrates also induced collagenase synthesis.
Sottile
J
J
Department of Biological Sciences, State University of New York, Albany 12222.
Mann
D M
DM
Diemer
V
V
Millis
A J
AJ
D002478:Cells, Cultured; D003470:Culture Media; D003513:Cycloheximide / Q000494:pharmacology; D004790:Enzyme Induction; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000201:enzymology*; D006801:Humans; D003012:Microbial Collagenase / Q000096:biosynthesis / Q000235:genetics*; D008970:Molecular Weight
D003470:Culture Media; D003513:Cycloheximide; D003012:Microbial Collagenase