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PMID: 6341993 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cultured human fibroblasts synthesize and secrete thrombospondin and incorporate it into extracellular matrix.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 80 ·No. 4 ·1983-02-00 ·Pages 998-1002

Jaffe EA, Ruggiero JT, Leung LK, Doyle MJ, McKeown-Longo PJ, Mosher DF

Abstract

Thrombospondin, a major glycoprotein released from alpha granules of thrombin-stimulated platelets, is a disulfide-bonded trimer of 160-kilodalton subunits. Cultured human foreskin and fetal lung fibroblasts secreted thrombospondin (determined by enzyme-linked immunosorbent assay) into the culture medium in a time-dependent manner (15.7 and 5.8 micrograms per 10(6) cells per 24 hr, respectively); secretion was blocked by cycloheximide. [3H]Thrombospondin was isolated from [3H]leucine-labeled fibroblast postculture medium and from cell layers with rabbit polyclonal or mouse monoclonal anti-thrombospondin coupled to staphylococcal protein A-Sepharose. The immunologically isolated [3H]thrombospondin migrated in NaDodSO4/polyacrylamide gels with purified marker platelet thrombospondin both with and without reduction. Immunofluorescence microscopy using rabbit polyclonal and mouse monoclonal anti-thrombospondin antibodies localized thrombospondin to the fibrillar extracellular matrix surrounding the cells. Thus, cultured human fibroblasts secrete thrombospondin and incorporate it into the extracellular matrix.

MeSH Terms
Cells, Cultured Extracellular Space/physiology Fibroblasts/metabolism Fluorescent Antibody Technique Glycoproteins/metabolism Humans Kinetics Male Thrombospondins
Chemicals
Glycoproteins Thrombospondins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Jaffe E A
Ruggiero J T
Leung L K
Doyle M J
McKeown-Longo P J
Mosher D F
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.80.4.998
Published
1983-02-00
Pages
998-1002
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC393515
Subset
IM
Grants
NHLBI NIH HHS · HL-18828 · United States
NHLBI NIH HHS · HL-21644 · United States
NHLBI NIH HHS · HL-24885 · United States
External Links
PubMed source DOI: 10.1073/pnas.80.4.998 PMC Full Text View journal details
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