title: Quantification of collagen contraction in three-dimensional cell culture.
abstract: Many different cell types including fibroblasts, smooth muscle cells, endothelial cells, and cancer cells exert traction forces on the fibrous components of the extracellular matrix. This can be observed as matrix contraction both macro- and microscopically in three-dimensional (3D) tissues models such as collagen type I gels. The quantification of local contraction at the micron scale, including its directionality and speed, in correlation with other parameters such as cell invasion, local protein or gene expression, can provide useful information to study wound healing, organism development, and cancer metastasis. In this article, we present a set of tools to quantify the flow dynamics of collagen contraction, induced by cells migrating out of a multicellular cancer spheroid into a three-dimensional (3D) collagen matrix. We adapted a pseudo-speckle technique that can be applied to bright-field and fluorescent microscopy time series. The image analysis presented here is based on an in-house written software developed in the Matlab (Mathworks) programming environment. The analysis program is freely available from GitHub following the link: http://dx.doi.org/10.5281/zenodo.10116. This tool provides an automatized technique to measure collagen contraction that can be utilized in different 3D cellular systems.
authors: Centre de Recherche, Institut Curie, Paris Cedex 05, France; Centre National de la Recherche Scientifique, Paris Cedex 05, France; UPMC University Paris VI, Paris, France.
authors: Katarzyna S
authors: KS
authors: Kopanska
authors: Centre de Recherche, Institut Curie, Paris Cedex 05, France; Centre National de la Recherche Scientifique, Paris Cedex 05, France; UPMC University Paris VI, Paris, France.
authors: Matthias
authors: M
authors: Bussonnier
authors: Centre de Recherche, Institut Curie, Paris Cedex 05, France; Centre National de la Recherche Scientifique, Paris Cedex 05, France.
authors: Sara
authors: S
authors: Geraldo
authors: Centre de Recherche, Institut Curie, Paris Cedex 05, France; Centre National de la Recherche Scientifique, Paris Cedex 05, France.
authors: Anthony
authors: A
authors: Simon
authors: Centre de Recherche, Institut Curie, Paris Cedex 05, France; Centre National de la Recherche Scientifique, Paris Cedex 05, France.
authors: Danijela
authors: D
authors: Vignjevic
authors: Centre de Recherche, Institut Curie, Paris Cedex 05, France; Centre National de la Recherche Scientifique, Paris Cedex 05, France; UPMC University Paris VI, Paris, France.
authors: Timo
authors: T
authors: Betz
mesh_terms: D000465:Algorithms; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D003094:Collagen / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D051379:Mice; D012984:Software; D014584:User-Computer Interface
chemical_list: D049452:Green Fluorescent Proteins; D003094:Collagen
keywords: Collagen; Contraction; Image analysis; Pseudo-speckle; Three-dimensional cell cultures