Slicing and Culturing Pig Hearts under Physiological Conditions

Qinghui Ou; Riham  R.E. Abouleisa; Xian-Liang Tang; Hamzah  R. Juhardeen; Moustafa  H. Meki; Jessica  M. Miller; Guruprasad Giridharan; Ayman El-Baz; Roberto Bolli; Tamer  M.A. Mohamed

doi:10.3791/60913

Method Article

Slicing and Culturing Pig Hearts under Physiological Conditions

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DOI:

10.3791/60913

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March 20th, 2020

Qinghui Ou^*¹ , Riham R.E. Abouleisa^*¹ , Xian-Liang Tang¹ , Hamzah R. Juhardeen¹ , Moustafa H. Meki² , Jessica M. Miller² , Guruprasad Giridharan² , Ayman El-Baz² , Roberto Bolli¹ , Tamer M.A. Mohamed¹^,³^,⁴^,⁵^,⁶

¹Institute of Molecular Cardiology, Department of Medicine, University of Louisville, ²Department of Bioengineering, University of Louisville, ³Diabetes and Obesity Center, Department of Medicine, University of Louisville, ⁴Department of Pharmacology and Toxicology, University of Louisville, ⁵Institute of Cardiovascular Sciences, University of Manchester, ⁶Faculty of Pharmacy, Zagazig University

^* These authors contributed equally

Summary

This protocol describes how to slice and culture heart tissue under physiological conditions for 6 days. This culture system could be used as a platform for testing the efficacy of novel heart failure therapeutics as well as reliable testing of acute cardiotoxicity in a 3D heart model.

Abstract

Many novel drugs fail in clinical studies due to cardiotoxic side effects as the currently available in vitro assays and in vivo animal models poorly predict human cardiac liabilities, posing a multi-billion-dollar burden on the pharmaceutical industry. Hence, there is a worldwide unmet medical need for better approaches to identify drug cardiotoxicity before undertaking costly and time consuming 'first in man' trials. Currently, only immature cardiac cells (human induced pluripotent stem cell-derived cardiomyocytes [hiPSC-CMs]) are used to test therapeutic efficiency and drug toxicity as they are the only human cardiac cells that can be cultured for prolonged periods required to test drug efficacy and toxicity. However, a single cell type cannot replicate the phenotype of the complex 3D heart tissue which is formed of multiple cell types. Importantly, the effect of drugs needs to be tested on adult cardiomyocytes, which have different characteristics and toxicity responses compared to immature hiPSC-CMs. Culturing human heart slices is a promising model of intact human myocardium. This technology provides access to a complete multicellular system that mimics the human heart tissue and reflects the physiological or pathological conditions of the human myocardium. Recently, through optimization of the culture media components and the culture conditions to include continuous electrical stimulation at 1.2 Hz and intermittent oxygenation of the culture medium, we developed a new culture system setup that preserves viability and functionality of human and pig heart slices for 6 days in culture. In the current protocol, we are detailing the method for slicing and culturing pig heart as an example. The same protocol is used to culture slices from human, dog, sheep, or cat hearts. This culture system has the potential to become a powerful predictive human in situ model for acute cardiotoxicity testing that closes the gap between preclinical and clinical testing results.

Introduction

Drug induced cardiotoxicity is a major cause of market withdrawal¹. In the last decade of the 20^th century, eight non-cardiovascular drugs were withdrawn from the market as they resulted in sudden death due to ventricular arrhythmias². In addition, several anti-cancer therapies (while in many cases effective) can lead to several cardiotoxic effects including cardiomyopathy and arrhythmias. For example, both traditional (e.g., anthracyclines and radiation) and targeted (e.g., trastuzumab) breast cancer therapies can result in cardiovascular complications in a subset of patients³. A close....