Quantitative analysis of 3D extracellular matrix remodelling by pancreatic stellate cells.

Benjamin K Robinson,Ernesto Cortes,Armando Del Río Hernández,Muge Sarper,Alistair J Rice

doi:10.1242/bio.017632

Abstract

ABSTRACTExtracellular matrix (ECM) remodelling is integral to numerous physiological and pathological processes in biology, such as embryogenesis, wound healing, fibrosis and cancer. Until recently, most cellular studies have been conducted on 2D environments where mechanical cues significantly differ from physiologically relevant 3D environments, impacting cellular behaviour and masking the interpretation of cellular function in health and disease. We present an integrated methodology where cell-ECM interactions can be investigated in 3D environments via ECM remodelling. Monitoring and quantification of collagen-I structure in remodelled matrices, through designated algorithms, show that 3D matrices can be used to correlate remodelling with increased ECM stiffness observed in fibrosis. Pancreatic stellate cells (PSCs) are the key effectors of the stromal fibrosis associated to pancreatic cancer. We use PSCs to implement our methodology and demonstrate that PSC matrix remodelling capabilities depend on their contractile machinery and β1 integrin-mediated cell-ECM attachment.

Highlights

3D remodelling of the extracellular matrix (ECM), which involves changes in ECM rigidity and organisation, is integral to several biological processes, such as wound healing (Darby et al, 2014; Reinke and Sorg, 2012), fibrosis (Duscher et al, 2014; Ho et al, 2014), and embryogenesis, where mechanical forces dictate tissue organisation (Krieg et al, 2008)
In pancreatic ductal adenocarcinoma (PDAC), the strong fibrosis in the stromal region around the tumour is mediated via ECM remodelling and orchestrated by pancreatic stellate cells (PSCs) (Apte et al, 2011; Apte and Wilson, 2012; Olsen et al, 2011)
second harmonic generation (SHG) is a microscopy technique that is highly responsive to fibrillar collagen (Chen et al, 2012)

Summary

Introduction

Cellular and Molecular Biomechanics Laboratory, Department of Bioengineering, Faculty of Engineering, Imperial College London, South Kensington Campus, London SW7 2AZ, UK. Received 15 February 2016; Accepted 23 April 2016 cancer cell invasion (Goetz et al, 2011). Assessing these quantitative changes in the ECM will provide a better understanding of the remodelling processes

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