Extracellular matrix contraction by cultured mesangial cells: an assay system for mesangial cell-matrix interaction.
To investigate the interaction between glomerular mesangial cells (MCs) and extracellular matrix (ECM), we have developed a quantitative assay using a floating culture of MC-populated collagen gel matrix (MCGM). Since MCs can contract such a matrix consisting of type I collagen, MCGM exhibited marked contraction. The apparent cause was tensional interaction between MCs and collagen fibers, because: (1) MCs in the gel attached tightly to surrounding collagen fibers; (2) collagen fibrils surrounding MCs were ordered in a radial array; (3) collagen fibers aggregated around MCs in the contracted gel, but not in an uncontracted gel; (4) cytochalasin B, an actin polymerization blocker, inhibited gel contraction in a dose-dependent manner. We found that this interaction was modulated by some factors. Serum in the medium stimulated the contraction of MCGM. The degree of MCGM contraction was proportional to the number of MCs embedded above 2.3 x 10(4) cells/ml of gel. In MCGM containing a basement membrane-type gel matrix (BGM), gel contraction was increasingly inhibited as the content of BGM rose. Our method is useful for elucidating physiological and pathological interactions between MCs and ECM.
Kitamura
M
M
Department of Internal Medicine, Jikei University School of Medicine, Tokyo, Japan.
Maruyama
N
N
Yoshida
H
H
Nagasawa
R
R
Mitarai
T
T
Sakai
O
O
D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D001485:Basement Membrane / Q000648:ultrastructure; D002448:Cell Adhesion; D002478:Cells, Cultured; D003094:Collagen; D003571:Cytochalasin B / Q000494:pharmacology; D004338:Drug Combinations; D005109:Extracellular Matrix / Q000737:chemistry / Q000648:ultrastructure*; D005920:Glomerular Mesangium / Q000166:cytology* / Q000187:drug effects; D066298:In Vitro Techniques; D007797:Laminin; D051379:Mice; D008807:Mice, Inbred BALB C; D008855:Microscopy, Electron, Scanning; D011509:Proteoglycans; D051381:Rats; D011919:Rats, Inbred Strains
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D000804:Angiotensin II; C059485:matrigel; D003571:Cytochalasin B; D003094:Collagen