scholarly journals Effect of cell-cell and cell-matrix interactions on the response of fibroblasts to epidermal growth factor in vitro. Expression of collagen type I, collagenase, stromelysin and tissue inhibitor of metalloproteinases

1992 ◽  
Vol 285 (1) ◽  
pp. 215-221 ◽  
Author(s):  
A C Colige ◽  
C A Lambert ◽  
B V Nusgens ◽  
C M Lapière
Keyword(s):  
Growth Factor ◽  
State Level ◽  
Collagen Type I ◽  
Culture Conditions ◽  
Type I ◽  
Cell Matrix ◽  
Epidermal Growth ◽  
Cell Matrix Interactions ◽  
Cell Cell

Investigations of the effect of epidermal growth factor (EGF) on the expression of four genes involved in the turnover of the extracellular matrix, collagen type I, collagenase, stromelysin and tissue inhibitor of metalloproteinases (TIMP) were performed on four strains of skin fibroblasts in vitro. Addition of EGF to subconfluent cultures for increasing periods of time up to 5 days induced an inhibition of procollagen alpha 1(I) mRNA and a strong stimulation of collagenase (100-fold) and stromelysin (1000-fold) mRNAs, whereas the mRNA of TIMP was increased to a lesser extent (5-fold). After a 40 h pulse with EGF, these effects persisted for 24-48 h after withdrawal of the growth factor and slowly diminished thereafter to attain control values after several days. By culturing fibroblasts for increasing periods of time, different levels of confluence were obtained allowing for the deposition of an extracellular biomatrix. The steady-state level of collagenase and stromelysin mRNAs were profoundly depressed in confluent as against non-confluent cultures, whereas no major change for TIMP and procollagen alpha 1(I) mRNAs was observed. Upon treatment of these cultures with EGF for 48h, the steady-state level of collagenase, stromelysin and TIMP increased, whereas procollagen alpha 1(I) mRNA was slightly reduced. These modifications were, at least in part, dependent upon a regulation of the transcription rate, as suggested from run-off experiments. Similar states of confluence were obtained by seeding cells at increasing densities in short-term cultures in which cell-cell contact predominated. In such culture conditions, the collagenase and stromelysin mRNAs were enhanced in high as compared to low density cultures. The response to EGF was progressively decreased for collagenase, stromelysin and, to a lesser extent, TIMP mRNAs at most densities and a complete lack of response to EGF at the highest cell density was observed. Under all culture conditions the modulation of collagenase mRNA was paralleled by similar modifications of enzyme activity. These results emphasize the importance of the cell-cell contacts and cell-matrix interactions in the expression of the genes coding for metalloproteinases or their inhibitor and their modulation by growth factors.

scholarly journals Changes in integrin expression during chondrogenesis in vitro: an immunomorphological study.

1995 ◽  
Vol 43 (10) ◽  
pp. 1061-1069 ◽  
Author(s):  
M Shakibaei ◽  
B Zimmermann ◽  
H J Merker
Keyword(s):  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Integrin Expression ◽  
Collagen Formation ◽  
Cell Matrix ◽  
Cell Type Specific ◽  
Immunomorphological Study ◽  
Cell Matrix Interactions

Integrins are receptors composed of ligand-specific alpha-chains and cell type-specific beta-chains which are involved in cell-cell and cell-matrix interactions. The distribution of alpha 1- and alpha 3-integrins as well as collagen Types I and II, was investigated by immunofluorescence and immunoelectron microscopy during chondrogenesis in organ culture after various culture periods. Mesenchymal cells from limb buds of Day 12 mouse embryos were grown at high density. Within the first 2 days of the culture period, only alpha 1-integrin could be detected. Formation of cartilage-specific matrix on Day 3 was accompanied by the occurrence of alpha 3-integrin. On Day 7, alpha 3 was present only in cartilage nodules, whereas alpha 1 was strongly expressed in the perichondrium and was more or less homogeneously distributed in the surrounding mesenchyme. On Day 14, alpha 1-integrin was again detectable in cartilage. We suggest that the change in collagen formation from Type I to Type II during chondrogenesis is accompanied by a change in integrin expression from alpha 1 to alpha 3. Conversely, dedifferentiation of chondrocytes in aging cartilage is accompanied by the occurrence of collagen Type I and alpha 1-integrin. Therefore, a strict correlation between the collagen type synthesized by the cells and the appropriate receptor presented by the cells is suggested.

Intestinal transformation results in transforming growth factor-beta-dependent alteration in tumor cell-cell matrix interactions

Surgery ◽  
2003 ◽  
Vol 133 (5) ◽  
pp. 568-579 ◽  
Author(s):  
David H. Berger ◽  
Christine A. O'Mahony ◽  
Hongmiao Sheng ◽  
Jinyi Shao ◽  
Daniel Albo ◽  
...  
Keyword(s):  
Growth Factor ◽  
Tumor Cell ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Growth Factor Beta ◽  
Cell Matrix Interactions ◽  
Cell Cell

scholarly journals Collagen Type I Containing Hybrid Hydrogel Enhances Cardiomyocyte Maturation in a 3D Cardiac Model

Polymers ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 687 ◽  
Author(s):  
Sam G. Edalat ◽  
Yongjun Jang ◽  
Jongseong Kim ◽  
Yongdoo Park
Keyword(s):  
Growth Factor ◽  
Collagen Type ◽  
Transforming Growth Factor ◽  
Embryonic Stem ◽  
Collagen Type I ◽  
Transforming Growth Factor Β1 ◽  
Type I ◽  
Hybrid Hydrogel ◽  
Cardiac Model

In vitro maturation of cardiomyocytes in 3D is essential for the development of viable cardiac models for therapeutic and developmental studies. The method by which cardiomyocytes undergoes maturation has significant implications for understanding cardiomyocytes biology. The regulation of the extracellular matrix (ECM) by changing the composition and stiffness is quintessential for engineering a suitable environment for cardiomyocytes maturation. In this paper, we demonstrate that collagen type I, a component of the ECM, plays a crucial role in the maturation of cardiomyocytes. To this end, embryonic stem-cell derived cardiomyocytes were incorporated into Matrigel-based hydrogels with varying collagen type I concentrations of 0 mg, 3 mg, and 6 mg. Each hydrogel was analyzed by measuring the degree of stiffness, the expression levels of MLC2v, TBX18, and pre-miR-21, and the size of the hydrogels. It was shown that among the hydrogel variants, the Matrigel-based hydrogel with 3 mg of collagen type I facilitates cardiomyocyte maturation by increasing MLC2v expression. The treatment of transforming growth factor β1 (TGF-β1) or fibroblast growth factor 4 (FGF-4) on the hydrogels further enhanced the MLC2v expression and thereby cardiomyocyte maturation.

scholarly journals Peptide gels of fully-defined composition and mechanics for probing cell-cell and cell-matrix interactions in vitro

2020 ◽  
Vol 85-86 ◽  
pp. 15-33 ◽  
Author(s):  
J.C. Ashworth ◽  
J.L. Thompson ◽  
J.R. James ◽  
C.E. Slater ◽  
S. Pijuan-Galitó ◽  
...  
Keyword(s):  
Cell Matrix ◽  
Matrix Interactions ◽  
Cell Matrix Interactions ◽  
Cell Cell

scholarly journals Organotypic Cultures of Intervertebral Disc Cells: Responses to Growth Factors and Signaling Pathways Involved

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Harris Pratsinis ◽  
Dimitris Kletsas
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Intervertebral Disc ◽  
Dna Synthesis ◽  
Signaling Pathways ◽  
Collagen Type I ◽  
Cell Physiology ◽  
Type I ◽  
Collagen Gels

Intervertebral disc (IVD) degeneration is strongly associated with low back pain, a major cause of disability worldwide. An in-depth understanding of IVD cell physiology is required for the design of novel regenerative therapies. Accordingly, aim of this work was the study of IVD cell responses to mitogenic growth factors in a three-dimensional (3D) organotypic milieu, comprising characteristic molecules of IVD’s extracellular matrix. In particular, annulus fibrosus (AF) cells were cultured inside collagen type-I gels, while nucleus pulposus (NP) cells in chondroitin sulfate A (CSA) supplemented collagen gels, and the effects of Platelet-Derived Growth Factor (PDGF), basic Fibroblast Growth Factor (bFGF), and Insulin-Like Growth Factor-I (IGF-I) were assessed. All three growth factors stimulated DNA synthesis in both AF and NP 3D cell cultures, with potencies similar to those observed previously in monolayers. CSA supplementation inhibited basal DNA synthesis rates, without affecting the response to growth factors. ERK and Akt were found to be phosphorylated following growth factor stimulation. Blockade of these two signaling pathways using pharmacologic inhibitors significantly, though not completely, inhibited growth factor-induced DNA synthesis. The proposed culture systems may prove useful for further in vitro studies aiming at future interventions for IVD regeneration.

Rapid generation of collagen-based microtissues to study cell–matrix interactions

TECHNOLOGY ◽  
2016 ◽  
Vol 04 (02) ◽  
pp. 80-87 ◽  
Author(s):  
Marie-Elena Brett ◽  
Alexandra L. Crampton ◽  
David K. Wood
Keyword(s):  
Large Scale ◽  
Culture Conditions ◽  
Encapsulated Cells ◽  
Cell Contractility ◽  
Tissue Constructs ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Rapid Generation ◽  
Cell Matrix Interactions

The objective of this study was to create a method for studying cell–matrix interactions in a physiologically relevant 3D protein-based tissue construct that could be scaled up to perform large-scale screens, study cell–matrix interactions on a population basis, or be remodeled by cells to build larger tissues. We have developed an easy-to-use method to miniaturize protein-based tissue constructs that maintains the 3D in vitro environment, while alleviating several obstacles associated with larger avascular tissue constructs. In this study, we demonstrate that (i) cells can interact with the 3D environment both while encapsulated or while interacting only with the surface of the microtissues, (ii) encapsulated cells are highly viable and, for the first time, (iii) microtissues on this size scale (~200 μm) can be used to quantify cell contractility. This versatile platform should facilitate large-scale screens in 3D in vitro culture conditions for drug development and high throughput mechanistic biology.

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