title: Living cardiac tissue slices: an organotypic pseudo two-dimensional model for cardiac biophysics research.
abstract: Living cardiac tissue slices, a pseudo two-dimensional (2D) preparation, have received less attention than isolated single cells, cell cultures, or Langendorff-perfused hearts in cardiac biophysics research. This is, in part, due to difficulties associated with sectioning cardiac tissue to obtain live slices. With moderate complexity, native cell-types, and well-preserved cell-cell electrical and mechanical interconnections, cardiac tissue slices have several advantages for studying cardiac electrophysiology. The trans-membrane potential (Vm) has, thus far, mainly been explored using multi-electrode arrays. Here, we combine tissue slices with optical mapping to monitor Vm and intracellular Ca(2+) concentration ([Ca(2+)]i). This combination opens up the possibility of studying the effects of experimental interventions upon action potential (AP) and calcium transient (CaT) dynamics in 2D, and with relatively high spatio-temporal resolution. As an intervention, we conducted proof-of-principle application of stretch. Mechanical stimulation of cardiac preparations is well-established for membrane patches, single cells and whole heart preparations. For cardiac tissue slices, it is possible to apply stretch perpendicular or parallel to the dominant orientation of cells, while keeping the preparation in a constant focal plane for fluorescent imaging of in-slice functional dynamics. Slice-to-slice comparison furthermore allows one to assess transmural differences in ventricular tissue responses to mechanical challenges. We developed and tested application of axial stretch to cardiac tissue slices, using a manually-controlled stretching device, and recorded Vm and [Ca(2+)]i by optical mapping before, during, and after application of stretch. Living cardiac tissue slices, exposed to axial stretch, show an initial shortening in both AP and CaT duration upon stretch application, followed in most cases by a gradual prolongation of AP and CaT duration during stretch maintained for up to 50 min. After release of sustained stretch, AP duration (APD) and CaT duration reverted to shorter values. Living cardiac tissue slices are a promising experimental model for the study of cardiac mechano-electric interactions. The methodology described here can be refined to achieve more accurate control over stretch amplitude and timing (e.g. using a computer-controlled motorised stage, or by synchronising electrical and mechanical events) and through monitoring of regional tissue deformation (e.g. by adding motion tracking).
authors.affiliation: Computer Science Department, University of Oxford, Wolfson Building, Parks Road, Oxford, OX1 3QD, UK. Electronic address: ken.wang@dtc.ox.ac.uk.
authors.forename: Ken
authors.initials: K
authors.lastname: Wang
authors.affiliation: Department of Pharmacology, University of Oxford, Mansfield Road, Oxford OX1 3QT, UK. Electronic address: derek.terrar@pharm.ox.ac.uk.
authors.forename: Derek
authors.initials: D
authors.lastname: Terrar
authors.affiliation: Computer Science Department, University of Oxford, Wolfson Building, Parks Road, Oxford, OX1 3QD, UK. Electronic address: david.gavaghan@cx.ox.ac.uk.
authors.forename: David J
authors.initials: DJ
authors.lastname: Gavaghan
authors.affiliation: Qatar Cardiovascular Research Center, Qatar Foundation Qatar Foundation, PO Box 5825, Doha, Qatar. Electronic address: rmuumin@qf.org.qa.
authors.forename: Razik
authors.initials: R
authors.lastname: Mu-U-Min
authors.affiliation: Computer Science Department, University of Oxford, Wolfson Building, Parks Road, Oxford, OX1 3QD, UK; Harefield Heart Science Centre, National Heart and Lung Institute, Imperial College London, Harefield, Middlesex, UB9 6JH, UK. Electronic address: p.kohl@imperial.ac.uk.
authors.forename: Peter
authors.initials: P
authors.lastname: Kohl
authors.affiliation: Qatar Cardiovascular Research Center, Qatar Foundation Qatar Foundation, PO Box 5825, Doha, Qatar; Harefield Heart Science Centre, National Heart and Lung Institute, Imperial College London, Harefield, Middlesex, UB9 6JH, UK. Electronic address: cbollensdorff@qf.org.qa.
authors.forename: Christian
authors.initials: C
authors.lastname: Bollensdorff
mesh_terms: D000200:Action Potentials / Q000502:physiology*; D000818:Animals; D020013:Calcium Signaling / Q000502:physiology*; D002478:Cells, Cultured; D056966:Excitation Contraction Coupling / Q000502:physiology*; D006329:Heart Conduction System / Q000502:physiology*; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D009924:Organ Culture Techniques / Q000379:methods*; D010812:Physical Stimulation / Q000379:methods; D011817:Rabbits; D013314:Stress, Mechanical
keywords: Axial stretch; Mechano-electric feedback; Multi-parametric; Optical mapping