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Abstract

Medicine

Preparation of Human Myocardial Tissue for Long-Term Cultivation

Published: June 2nd, 2022

DOI:

10.3791/63964

1Walter-Brendel-Centre of Experimental Medicine, University Hospital, LMU Munich, 2German Center for Cardiovascular Research (DZHK), Partner Site Munich, Munich Heart Alliance (MHA), 3Division of Experimental Cardiology, Department of Cardiology, Erasmus MC, University Medical Center Rotterdam, 4Institute of Cellular and Molecular Physiology, Friedrich-Alexander-Universität Erlangen-Nürnberg, 5Muscle Research Center Erlangen (MURCE), Friedrich-Alexander-Universität Erlangen-Nürnberg, 6Department of Cardiac Surgery, University Hospital, LMU Munich

Abstract

Cardiomyocyte cultivation has seen a vast number of developments, ranging from two-dimensional (2D) cell cultivation to iPSC derived organoids. In 2019, an ex vivo way to cultivate myocardial slices obtained from human heart samples was demonstrated, while approaching in vivo condition of myocardial contraction. These samples originate mostly from heart transplantations or left-ventricular assist device placements. Using a vibratome and a specially developed cultivation system, 300 µm thick slices are placed between a fixed and a spring wire, allowing for stable and reproducible cultivation for several weeks. During cultivation, the slices are continuously stimulated according to individual settings. Contractions can be displayed and recorded in real-time, and pharmacological agents can be readily applied. User-defined stimulation protocols can be scheduled and performed to assess vital contraction parameters like post-pause-potentiation, stimulation threshold, force-frequency relation, and refractory period. Furthermore, the system enables a variable pre- and afterload setting for a more physiological cultivation.

Here, we present a step-by-step guide on how to generate a successful long-term cultivation of human left ventricular myocardial slices, using a commercial biomimetic cultivation solution.

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Keywords Human Myocardial Tissue

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