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Related Experiment Videos

Thrombin stimulation of matrix fibronectin

M T Armstrong1, J W Fenton, T T Andersen

  • 1Department of Molecular and Cellular Biology, University of California, Davis 95616-8755, USA.

Journal of Cellular Physiology
|January 1, 1996
PubMed
Summary

Growth factors stimulate cell proliferation by inducing fibronectin matrix production in 3D fibroblast cultures. This matrix is essential for mitogenic signaling in mesenchymal tissues, unlike in 2D cultures.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Extracellular Matrix Research

Background:

  • Thrombin and trypsin are known to stimulate cell proliferation.
  • The role of the extracellular matrix (ECM) in cell signaling is an area of active research.

Purpose of the Study:

  • To investigate the role of fibronectin matrix production in growth factor-induced cell proliferation.
  • To determine if matrix production is a prerequisite for mitogenic signaling in mesenchymal tissues.

Main Methods:

  • Culturing embryonic fibroblasts as 3D tissue-like aggregates and 2D monolayers.
  • Stimulating cultures with trypsin, thrombin, and specific peptide analogues (SFLLRN, SFLLRNPNDKYEPF).
  • Assessing fibronectin matrix elaboration, cell proliferation, and the effect of RGD peptides.

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Main Results:

  • SFLLRN and thrombin stimulated fibronectin-rich ECM production and cell proliferation in 3D fibroblast aggregates.
  • Matrix production was essential for SFLLRN-induced proliferation, as shown by inhibition with RGD peptides.
  • Fibronectin matrix preparations stimulated proliferation in growth factor-free medium.
  • In contrast, thrombin and trypsin induced proliferation in 2D monolayer cultures without stimulating matrix production.

Conclusions:

  • Fibronectin matrix elaboration is a necessary event for mitogenic signaling in mesenchymal tissues.
  • A tight coupling exists between matrix production (matrigenic activity) and cell proliferation (mitogenic activity) in 3D cultures.
  • This coupling is absent in 2D monolayer cultures, highlighting the importance of the tissue microenvironment.