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Modulation of extracellular‐matrix synthesized by cultured stromal cells from normal human breast tissue by epidermal growth factor
Author(s) -
Baley Patricia,
Lützelschwab Irene,
ScottBurden Timothy,
Küng Willy,
Eppenberger Urs
Publication year - 1990
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240430203
Subject(s) - epidermal growth factor , extracellular matrix , tenascin , fibronectin , stromal cell , laminin , matrix (chemical analysis) , growth factor , cell culture , microbiology and biotechnology , tissue culture , biology , chemistry , biochemistry , receptor , cancer research , in vitro , genetics , chromatography
A routine, reproducible procedure was developed for the preparation and characterization of stromal cells from normal human breast tissue obtained by reduction mammaplasty. Isolates (n = 15) all exhibited enhanced rates of proliferation, even in the presence of 20% fetal calf serum, when exposed to epidermal growth factor or transforming growth factor α (both 10 −8 M). Cellular responsiveness to these growth factors was consistent with expression of specific surface receptors for epidermal growth factor (∼10 4 /cell). In cultures, stromal cells elaborated an extensive, crosslinked, insoluble extracellular matrix which remained firmly associated with the plastic surface of tissue culture ware upon lysis of cells. The insoluble matrix material was analyzed using enzymatic digestion procedures following incorporation of radio‐labelled precursors into macromolecular material prior to lysis and preparation. The relative proportion of glycoconjugate (glycopeptides and proteoglycans) and collage‐nous material present in matrix material was ∼45% and ∼55%, respectively, and this was modulated by inclusion of epidermal growth factor into culture medium to ∼60% and ∼40%, respectively. Under similar culture conditions stromal cells synthesized twice as much hyaluronate as was produced by control cultures. By use of specific antibody preparations we identified at least four species of glycopeptide present in stromal matrices (namely, fibronectin, laminin, tenascin, and thrombospon‐din) as well as three types of collagen (types I, III, and IV). The rapid and reproducible procedure for the preparation of radiolabelled insoluble matrix material from normal human breast tissue allows for the study of cellular interaction involving extracellular matrix turnover and degradation.