Collagenase production by early and late passage cultures of human fibroblasts

doi:10.1016/0531-5565(89)90060-0

Experimental Gerontology

Volume 24, Issues 5–6, 1989, Pages 559-575

https://doi.org/10.1016/0531-5565(89)90060-0 Get rights and content

Abstract

Comparison of the proteins secreted by early and late passage cell cultures of human fibroblasts revealed a high level of immunoreactive collagenase (Mr = 55,000 Da and 58,000 aDa) in the late passage cell culture conditioned medium. Both molecular weight species reacted with a monoclonal anticollagenase antibody and were apparently glycosylation varients of the same protein. The question of whether the apparent age-dependent differences in collagenase synthesis reflected changes in protein synthesis or secretion was addressed by assaying immunoreactive collagenase and collagenase mRNA. Immunofluorescence microscopy of cellular collagenase revealed that the percentage of collagenase positive cells ranged from 1 to 6% (early passage) to 35 to 46% (late passage) indicating that the late passage cells had higher basal levels of collagenase synthesis. Later passage cultures also secreted higher levels of immunoprecipitable collagenase into the culture medium and Northern analysis established that the basal levels of collagenase mRNA was also 10 times greater in late passage cells. High basal levels of collagenase were also observed in fibroblasts cultured from an in vivo aged donor and from donors with Werner's syndrome. Collagenase production was induced in both early and late passage cell cultures by exposure to fibroblast extracellular matrix, fibroblast conditioned media, polypeptide growth factors, or phorbol esters. The induced levels were always greater in the late passage cell cultures than in the early passage cell cultures.

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¹: Current Address: Department of Physiological Chemistry, University of Wisconsin Medical School, Madison, WI 53706.

²: Current Address: La Jolla Cancer Research Foundation, 10901 North Torrey Pines Road, La Jolla, CA 92037.

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Collagenase production by early and late passage cultures of human fibroblasts

Abstract

Cell

Invest. Dermatol.

J. Biol. Chem.

J. Biol. Chem.

Coll. Relat. Res.

Exp. Cell Res.

Coll. Relat. Res.

Exp. Cell Res.

Mech. Ageing Dev.

Cell

Mech. Ageing Dev.

Coll. Relat. Res.

Mech. Ageing Dev.

J. Biol. Chem.

J. Biol. Chem.

Collagenase is a major gene product of induced rabbit synovial fibroblasts

J. Cell Biol.

12-O-tetradecanoyl-phorbol-13-acetate (TPA) induction of the human collagenase gene is mediated by an inducible enhancer element located in the 5′-flanking region

Mol. Cell Biol.

Collagenase

Stimulation of in vitro human skin collagenase expression by platelet-derived growth factor

Diminished response of Werner's syndrome fibroblasts to growth factors PDGF and FGF

Science

Matrix influence on the tumor cell stimulation of fibroblast collagenase production

Autoregulation of collagenase production by a protein synthesized and secreted by synovial fibroblasts: Cellular mechanism for control of collagen degradation

Molecular cloning of human synovial cell collagenase and selection of a single gene from genomic DNA

J. Clin. Invest.