Porcine Organotypic Epicardial Slice Protocol: A Tool for the Study of Epicardium in Cardiovascular Research

Davide Maselli; Rolando S. Matos; Robert D. Johnson; Davide Martella; Valeria Caprettini; Ciro Chiappini; Patrizia Camelliti; Paola Campagnolo

doi:10.3389/fcvm.2022.920013

Porcine Organotypic Epicardial Slice Protocol: A Tool for the Study of Epicardium in Cardiovascular Research

Davide Maselli, Rolando S. Matos, Robert D. Johnson, Davide Martella, Valeria Caprettini, Ciro Chiappini, Patrizia Camelliti, Paola Campagnolo^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

Abstract

The epicardium has recently gained interest in the cardiovascular field due to its capacity to support heart regeneration after ischemic injury. Models to study the epicardium of large animals in vitro are limited and mainly based on epicardial cell isolation/differentiation from stem cells, followed by 2D cells culture. In this method paper, we describe the procedure to obtain and culture 3D organotypic heart slices presenting an intact epicardium, as a novel model to study the epicardial physiology and activation. Epicardial slices are obtained from porcine hearts using a high-precision vibratome and retain a healthy epicardial layer embedded in its native extracellular environment and connected with other cardiac cells (cardiomyocytes, fibroblasts, vascular cells etc.). Epicardial slices can be cultured for 72 h, providing an ideal model for studying the epicardium physiology or perform pharmacological interventions/gene therapy approaches. We also report on methods to assesses the viability and composition of the epicardial slices, and evaluate their architecture in 3D through tissue decoloration. Finally, we present a potential application for a nanomaterial-based gene transfer method for tracking of epicardial cells within the slice. Crucially, given the similarity in morphology and physiology of porcine heart with its human counterpart, our system provides a platform for translational research while providing a clinically relevant and ethical alternative to the use of small animals in this type of research.

Original language	English
Article number	920013
Journal	Frontiers in Cardiovascular Medicine
Volume	9
DOIs	https://doi.org/10.3389/fcvm.2022.920013
Publication status	Published - 18 Jul 2022

Keywords

epicardium
ex vivo model
myocardial infarction
organotypic culture model
regenerative medicine

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.3389/fcvm.2022.920013

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title = "Porcine Organotypic Epicardial Slice Protocol: A Tool for the Study of Epicardium in Cardiovascular Research",

abstract = "The epicardium has recently gained interest in the cardiovascular field due to its capacity to support heart regeneration after ischemic injury. Models to study the epicardium of large animals in vitro are limited and mainly based on epicardial cell isolation/differentiation from stem cells, followed by 2D cells culture. In this method paper, we describe the procedure to obtain and culture 3D organotypic heart slices presenting an intact epicardium, as a novel model to study the epicardial physiology and activation. Epicardial slices are obtained from porcine hearts using a high-precision vibratome and retain a healthy epicardial layer embedded in its native extracellular environment and connected with other cardiac cells (cardiomyocytes, fibroblasts, vascular cells etc.). Epicardial slices can be cultured for 72 h, providing an ideal model for studying the epicardium physiology or perform pharmacological interventions/gene therapy approaches. We also report on methods to assesses the viability and composition of the epicardial slices, and evaluate their architecture in 3D through tissue decoloration. Finally, we present a potential application for a nanomaterial-based gene transfer method for tracking of epicardial cells within the slice. Crucially, given the similarity in morphology and physiology of porcine heart with its human counterpart, our system provides a platform for translational research while providing a clinically relevant and ethical alternative to the use of small animals in this type of research.",

keywords = "epicardium, ex vivo model, myocardial infarction, organotypic culture model, regenerative medicine",

author = "Davide Maselli and Matos, {Rolando S.} and Johnson, {Robert D.} and Davide Martella and Valeria Caprettini and Ciro Chiappini and Patrizia Camelliti and Paola Campagnolo",

note = "Funding Information: This work PCamp and DMas were supported by the National Centre for the Replacement, Refinement & Reduction of Animals in Research (grant numbers: NC/R001006/1 and NC/T001216/1). RM is supported by the Doctoral College studentship award (University of Surrey) and CC, DMar, and VC by the European Research Council (grant reference: StG EnBioN 759577). PCame and RJ acknowledge support from the British Heart Foundation (grant number: FS/17/33/32931), the Royal Society (RSG/R1/180198) and the HEIF Strategic Fund. Funding Information: We thank Drs. L. Dixton, A. Reis, and M. Henstock from the Pirbright Institute (Pirbright, UK) and the personnel at Newman's Abattoir (Farnborough, UK) for their support in procuring the animal tissues. Prof. John McVey and the Department of Biochemical Sciences at the University of Surrey, especially the technical team, for their continuing support. Publisher Copyright: Copyright {\textcopyright} 2022 Maselli, Matos, Johnson, Martella, Caprettini, Chiappini, Camelliti and Campagnolo.",

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doi = "10.3389/fcvm.2022.920013",

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T1 - Porcine Organotypic Epicardial Slice Protocol

T2 - A Tool for the Study of Epicardium in Cardiovascular Research

AU - Maselli, Davide

AU - Matos, Rolando S.

AU - Johnson, Robert D.

AU - Martella, Davide

AU - Caprettini, Valeria

AU - Chiappini, Ciro

AU - Camelliti, Patrizia

AU - Campagnolo, Paola

N1 - Funding Information: This work PCamp and DMas were supported by the National Centre for the Replacement, Refinement & Reduction of Animals in Research (grant numbers: NC/R001006/1 and NC/T001216/1). RM is supported by the Doctoral College studentship award (University of Surrey) and CC, DMar, and VC by the European Research Council (grant reference: StG EnBioN 759577). PCame and RJ acknowledge support from the British Heart Foundation (grant number: FS/17/33/32931), the Royal Society (RSG/R1/180198) and the HEIF Strategic Fund. Funding Information: We thank Drs. L. Dixton, A. Reis, and M. Henstock from the Pirbright Institute (Pirbright, UK) and the personnel at Newman's Abattoir (Farnborough, UK) for their support in procuring the animal tissues. Prof. John McVey and the Department of Biochemical Sciences at the University of Surrey, especially the technical team, for their continuing support. Publisher Copyright: Copyright © 2022 Maselli, Matos, Johnson, Martella, Caprettini, Chiappini, Camelliti and Campagnolo.

PY - 2022/7/18

Y1 - 2022/7/18

N2 - The epicardium has recently gained interest in the cardiovascular field due to its capacity to support heart regeneration after ischemic injury. Models to study the epicardium of large animals in vitro are limited and mainly based on epicardial cell isolation/differentiation from stem cells, followed by 2D cells culture. In this method paper, we describe the procedure to obtain and culture 3D organotypic heart slices presenting an intact epicardium, as a novel model to study the epicardial physiology and activation. Epicardial slices are obtained from porcine hearts using a high-precision vibratome and retain a healthy epicardial layer embedded in its native extracellular environment and connected with other cardiac cells (cardiomyocytes, fibroblasts, vascular cells etc.). Epicardial slices can be cultured for 72 h, providing an ideal model for studying the epicardium physiology or perform pharmacological interventions/gene therapy approaches. We also report on methods to assesses the viability and composition of the epicardial slices, and evaluate their architecture in 3D through tissue decoloration. Finally, we present a potential application for a nanomaterial-based gene transfer method for tracking of epicardial cells within the slice. Crucially, given the similarity in morphology and physiology of porcine heart with its human counterpart, our system provides a platform for translational research while providing a clinically relevant and ethical alternative to the use of small animals in this type of research.

AB - The epicardium has recently gained interest in the cardiovascular field due to its capacity to support heart regeneration after ischemic injury. Models to study the epicardium of large animals in vitro are limited and mainly based on epicardial cell isolation/differentiation from stem cells, followed by 2D cells culture. In this method paper, we describe the procedure to obtain and culture 3D organotypic heart slices presenting an intact epicardium, as a novel model to study the epicardial physiology and activation. Epicardial slices are obtained from porcine hearts using a high-precision vibratome and retain a healthy epicardial layer embedded in its native extracellular environment and connected with other cardiac cells (cardiomyocytes, fibroblasts, vascular cells etc.). Epicardial slices can be cultured for 72 h, providing an ideal model for studying the epicardium physiology or perform pharmacological interventions/gene therapy approaches. We also report on methods to assesses the viability and composition of the epicardial slices, and evaluate their architecture in 3D through tissue decoloration. Finally, we present a potential application for a nanomaterial-based gene transfer method for tracking of epicardial cells within the slice. Crucially, given the similarity in morphology and physiology of porcine heart with its human counterpart, our system provides a platform for translational research while providing a clinically relevant and ethical alternative to the use of small animals in this type of research.

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KW - ex vivo model

KW - myocardial infarction

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KW - regenerative medicine

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Porcine Organotypic Epicardial Slice Protocol: A Tool for the Study of Epicardium in Cardiovascular Research

Abstract

Keywords

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