Preparation of Extracellular Matrices Produced by Cultured and Primary Fibroblasts

Curr Protoc Cell Biol. 2016 Jun 1:71:10.9.1-10.9.34. doi: 10.1002/cpcb.2.

Abstract

Fibroblasts secrete and organize extracellular matrix (ECM), which provides structural support for their adhesion, migration, and tissue organization, besides regulating cellular functions such as growth and survival. Cell-to-matrix interactions are vital for vertebrate development. Disorders in these processes have been associated with fibrosis, developmental malformations, cancer, and other diseases. This unit describes a method for preparing a three-dimensional matrix derived from fibroblastic cells; the matrix is three-dimensional, cell and debris free, and attached to a two-dimensional culture surface. Cell adhesion and spreading are normal on these matrices. This matrix can also be compressed into a two-dimensional matrix and solubilized to study the matrix biochemically. © 2016 by John Wiley & Sons, Inc.

Keywords: 3D cell culture; cell-derived extracellular matrices; desmoplasia; myofibroblasts; primary fibroblasts; tumor- or cancer-associated fibroblasts.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Actins / metabolism
  • Animals
  • Blotting, Western
  • Cattle
  • Cell Culture Techniques / methods*
  • Cell Separation
  • Cell Shape
  • Cells, Cultured
  • Extracellular Matrix / metabolism*
  • Fibroblasts / metabolism*
  • Humans
  • Mice
  • NIH 3T3 Cells
  • Phenotype
  • Serum Albumin, Bovine / metabolism
  • Solubility
  • Staining and Labeling

Substances

  • Actins
  • alpha-smooth muscle actin, mouse
  • Serum Albumin, Bovine