Comparable Decellularization of Fetal and Adult Cardiac Tissue Explants as 3D-like Platforms for In Vitro Studies

Ana C. Silva; Maria J. Oliveira; Todd C McDevitt; Mário A. Barbosa; Diana S. Nascimento; Perpétua Pinto-do-Ó

doi:10.3791/56924

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Method Article

Comparable Decellularization of Fetal and Adult Cardiac Tissue Explants as 3D-like Platforms for In Vitro Studies

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DOI:

10.3791/56924

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March 21st, 2019

Ana C. Silva¹^,²^,³^,⁴ , Maria J. Oliveira¹^,²^,⁵ , Todd C McDevitt⁴^,⁶ , Mário A. Barbosa¹^,²^,³ , Diana S. Nascimento¹^,² , Perpétua Pinto-do-Ó¹^,²^,³

¹i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, ²INEB - Instituto de Engenharia Biomédica, Universidade do Porto, ³Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, ⁴Gladstone Institute of Cardiovascular Disease, ⁵Faculty of Medicine, University of Porto, ⁶University of California San Francisco

Erratum Notice

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Summary

The cardiac extracellular matrix (ECM) is a complex network of molecules that orchestrate key processes in tissues and organs while enduring physiological remodeling throughout life. Standardized decellularization of fetal and adult hearts permits comparative experimental studies of both tissues in a 3D context by capturing native architecture and biomechanical properties.

Abstract

Current knowledge of extracellular matrix (ECM)-cell communication translates to large two-dimensional (2D) in vitro culture studies where ECM components are presented as a surface coating. These culture systems constitute a simplification of the complex nature of the tissue ECM that encompasses biochemical composition, structure, and mechanical properties. To better emulate the ECM-cell communication shaping the cardiac microenvironment, we developed a protocol that allows for the decellularization of the whole fetal heart and adult left ventricle tissue explants simultaneously for comparative studies. The protocol combines the use of a hypotonic buffer, a detergent of anionic surfactant properties, and DNase treatment without any requirement for specialized skills or equipment. The application of the same decellularization strategy across tissue samples from subjects of various age is an alternative approach to perform comparative studies. The present protocol allows the identification of unique structural differences across fetal and adult cardiac ECM mesh and biological cellular responses. Furthermore, the herein methodology demonstrates a broader application being successfully applied in other tissues and species with minor adjustments, such as in human intestine biopsies and mouse lung.

Introduction

The extracellular matrix (ECM) is a dynamic network of molecules that regulate important cellular processes, namely fate-decision, proliferation and differentiation¹^,². The investigation of cell-ECM interactions has been performed mainly in two-dimensional (2D) in vitro cultures coated with ECM components, which constitute a simplification of native ECM properties found in vivo. Decellularization generates acellular 3D-like ECM bioscaffolds that largely preserve the extracellular architecture and composition of native tissues and organs³^,⁴. In....