Preparation of extracellular matrices produced by cultured and primary fibroblasts

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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.

Original languageEnglish
Pages (from-to)10.9.1-10.9.34.
JournalCurrent Protocols in Cell Biology
Volume2016
DOIs
StatePublished - 2016

Keywords

  • 3D cell culture
  • Cell-derived extracellular matrices
  • Desmoplasia
  • Myofibroblasts
  • Primary fibroblasts
  • Tumor-or cancer-associated fibroblasts
  • Extracellular Matrix/metabolism
  • NIH 3T3 Cells
  • Cell Culture Techniques/methods
  • Actins/metabolism
  • Cell Separation
  • Humans
  • Cells, Cultured
  • Solubility
  • Serum Albumin, Bovine/metabolism
  • Blotting, Western
  • Phenotype
  • Animals
  • Cattle
  • Fibroblasts/metabolism
  • Cell Shape
  • Staining and Labeling
  • Mice

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