Engineering clinically-relevant human fibroblastic cell-derived extracellular matrices

Janusz Franco-Barraza, Kristopher S. Raghavan, Tiffany Luong, Edna Cukierman

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

20 Scopus citations

Abstract

Three-dimensional (3D) culturing models, replicating in vivo tissue microenvironments that incorporate native extracellular matrix (ECM), have revolutionized the cell biology field. Fibroblastic cells generate lattices of interstitial ECM proteins. Cell interactions with ECMs and with molecules sequestered/stored within these are crucial for tissue development and homeostasis maintenance. Hence, ECMs provide cells with biochemical and biomechanical cues to support and locally control cell function. Further, dynamic changes in ECMs, and in cell-ECM interactions, partake in growth, development, and temporary occurrences such as acute wound healing. Notably, dysregulation in ECMs and fibroblasts could be important triggers and modulators of pathological events such as developmental defects, and diseases associated with fibrosis and chronic inflammation such as cancer. Studying the type of fibroblastic cells producing these matrices and how alterations to these cells enable changes in ECMs are of paramount importance. This chapter provides a step-by-step method for producing multilayered (e.g., 3D) fibroblastic cell-derived matrices (fCDM). Methods also include means to assess ECM topography and other cellular traits, indicative of fibroblastic functional statuses, like naïve/normal vs. inflammatory and/or myofibroblastic. For these, protocols include indications for isolating normal and diseased fibroblasts (i.e., cancer-associated fibroblasts known as CAFs). Protocols also include means for conducting microscopy assessments, querying whether fibroblasts present with fCDM-dependent normal or CAF phenotypes. These are supported by discrete semi-quantitative digital imaging analyses, providing some imaging processing advice. Additionally, protocols include descriptions for effective fCDM decellularization, which renders cellular debris-free patho/physiological in vivo-like scaffolds, suitable as 3D substrates for subsequent cell culturing.

Original languageEnglish
Title of host publicationCell-derived Matrices - Part A
EditorsDavid Caballero, Subhas C. Kundu, Rui Luís Reis
PublisherAcademic Press Inc.
Pages109-160
Number of pages52
Volume156
ISBN (Print)9780128201725
DOIs
StatePublished - 2020

Publication series

NameMethods in cell biology
ISSN (Print)0091-679X

Keywords

  • Cancer-associated fibroblasts
  • Cell-derived extracellular matrix
  • Cell-matrix interactions
  • Extracellular matrix
  • Primary fibroblasts
  • Three-dimensional cell culture
  • Tissue microenvironment

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