FACS analysis of Col1α protein levels in primary fibroblasts

Noam Cohen, Neta Erez

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

Abstract

Chronic inflammatory diseases are often associated with organ fibrosis, a progressive condition in which excessive deposition of extracellular matrix (ECM), mainly composed of collagen I (Col I), is deposited by activated fibroblasts and severely impairs tissue architecture and function, eventually resulting in organ failure. Moreover, enhanced collagen deposition by activated fibroblasts and increased stiffness of the extracellular matrix were demonstrated to be associated with tumor progression and metastasis. In order to quantitatively analyze fibrotic activation of fibroblasts and collagen deposition, it is essential to assess collagen content. While various histological methods allow assessment of collagen in tissue sections (e.g., Masson trichrome and Sirius red), reliable measurement and quantification of collagen levels in vitro remain a challenge in the field. In this protocol, we utilize intracellular staining of Col1α and flow cytometry analysis to analyze collagen content in primary fibroblasts isolated from fresh single cell suspensions of metastases-bearing lungs.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages221-228
Number of pages8
DOIs
StatePublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1944
ISSN (Print)1064-3745

Keywords

  • Cancer-associated fibroblasts
  • Collagen
  • FACS
  • Lung metastases

Fingerprint

Dive into the research topics of 'FACS analysis of Col1α protein levels in primary fibroblasts'. Together they form a unique fingerprint.

Cite this