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Related Experiment Video

Updated: Jul 20, 2025

Organotypic Collagen I Assay: A Malleable Platform to Assess Cell Behaviour in a 3-Dimensional Context
10:45

Organotypic Collagen I Assay: A Malleable Platform to Assess Cell Behaviour in a 3-Dimensional Context

Published on: October 13, 2011

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A quantitative in vitro collagen uptake assay.

Sjors Maassen1,2, Harry M Warner1, Pieter Grijpstra1

  • 1Department of Molecular Immunology, Groningen Biomolecular Sciences and Biotechnology, University of Groningen, Groningen, the Netherlands.

Methodsx
|August 3, 2023
PubMed
Summary

This study presents a new method to quantify collagen uptake by cells, crucial for understanding tissue remodelling. The protocol uses fluorescently labeled collagen and accutase for easy cell detachment and analysis.

Keywords:
A quantitative in vitro collagen uptake assayCollagenExtracellular matrixMacrophages

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Tissue Engineering

Background:

  • Collagen remodelling is essential for development and homeostasis.
  • While collagen production is understood, its removal by cells is less clear.
  • Understanding collagen removal is key to many biological processes.

Purpose of the Study:

  • To develop a quantitative method for measuring collagen uptake by cells.
  • To provide a reliable protocol for studying collagen degradation and turnover.
  • To facilitate research into cellular mechanisms of collagen processing.

Main Methods:

  • Utilizes a collagen type I-FITC conjugate polymerized in cell culture wells.
  • Cells are cultured on the collagen matrix and can be treated with various agents.
  • Employs accutase for efficient cell detachment, followed by FITC signal quantification via microscopy or flow cytometry.

Main Results:

  • The protocol allows for easy and quantitative measurement of cellular collagen uptake.
  • Accutase effectively detaches cells, including strongly adherent types like macrophages.
  • The method is compatible with various downstream analyses, including molecular isolation.

Conclusions:

  • This protocol offers a straightforward approach to quantify collagen uptake in cellular models.
  • It simplifies the study of collagen removal mechanisms, essential for tissue homeostasis.
  • The method supports diverse downstream applications, advancing research in cell biology and regenerative medicine.