A physiologic three-dimensional cell culture system to investigate the role of decorin in matrix organisation and cell survival.
In vivo cells exist in a three-dimensional environment generated and maintained by multiple cell-cell and cell-matrix interactions. Proteoglycans, like decorin, affect these complex interactions. Thus, we sought to investigate the role of decorin in a three-dimensional environment where the matrix was generated over time by decorin-deficient fibroblasts in the presence of L-ascorbic acid 2-phosphate. The cells were viable and proliferated in response to FGF2. Decorin was incorporated in the matrix and caused a approximately 2 nm shift in the average diameter of the collagen fibrils, and the range and distribution of the fibrils became narrower and more uniform. Although there were no appreciable changes in collagen composition, we found that exogenous decorin induced the de novo synthesis of collagen I and V and cross-linked beta(I). In the early phases of the three-dimensional culture, decorin reduced apoptosis. However, following the establishment of a three-dimensional matrix, the cells did not require decorin for their survival.
Seidler
Daniela G
DG
Department of Physiological Chemistry and Pathobiochemistry, Waldeyerstr. 15, University Hospital Münster, University of Münster, D-48149 Münster, Germany. dgseidle@uni-muenster.de
Schaefer
Liliana
L
Robenek
Horst
H
Iozzo
Renato V
RV
Kresse
Hans
H
Schönherr
Elke
E
D000818:Animals; D017209:Apoptosis; D001205:Ascorbic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D053148:Caspase 3; D053181:Caspase 8; D020169:Caspases / Q000378:metabolism; D002450:Cell Communication; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry; D024042:Collagen Type I / Q000737:chemistry; D024062:Collagen Type V / Q000737:chemistry; D058575:Decorin; D016326:Extracellular Matrix Proteins; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D008854:Microscopy, Electron; D010434:Pepsin A / Q000737:chemistry; D010641:Phenotype; D011509:Proteoglycans / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*
D024042:Collagen Type I; D024062:Collagen Type V; C546878:DCN protein, human; C546879:Dcn protein, mouse; D058575:Decorin; D016326:Extracellular Matrix Proteins; D011509:Proteoglycans; C011669:ascorbate-2-phosphate; D003094:Collagen; C505407:CASP3 protein, human; C505530:CASP8 protein, human; C505409:Casp3 protein, mouse; C505526:Casp8 protein, mouse; D053148:Caspase 3; D053181:Caspase 8; D020169:Caspases; D010434:Pepsin A; D001205:Ascorbic Acid