TY  - JOUR
A1  - Knox, Ronald
A1  - Bruggemann, Andrea
A1  - Gossmann, Matthias
A1  - Thomas, Ulrich
A1  - Horváth, András
A1  - Dragicevic, Elena
A1  - Stoelzle-Feix, Sonja
A1  - Fertig, Niels
A1  - Jung, Alexander
A1  - Raman, Aravind Hariharan
A1  - Staat, Manfred
A1  - Linder, Peter
T1  - Combining physiological relevance and throughput for in vitro cardiac contractility measurement
T2  - Biophysical Journal
N2  - Despite increasing acceptance of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in safety pharmacology, controversy remains about the physiological relevance of existing in vitro models for their mechanical testing. We hypothesize that existing signs of immaturity of the cell models result from an improper mechanical environment. We cultured hiPSC-CMs in a 96-well format on hyperelastic silicone membranes imitating their native mechanical environment, resulting in physiological responses to compound stimuli.We validated cell responses on the FLEXcyte 96, with a set of reference compounds covering a broad range of cellular targets, including ion channel modulators, adrenergic receptor modulators and kinase inhibitors. Acute (10 - 30 min) and chronic (up to 7 days) effects were investigated. Furthermore, the measurements were complemented with electromechanical models based on electrophysiological recordings of the used cell types.hiPSC-CMs were cultured on freely-swinging, ultra-thin and hyperelastic silicone membranes. The weight of the cell culture medium deflects the membranes downwards. Rhythmic contraction of the hiPSC-CMs resulted in dynamic deflection changes which were quantified by capacitive distance sensing. The cells were cultured for 7 days prior to compound addition. Acute measurements were conducted 10-30 minutes after compound addition in standard culture medium. For chronic treatment, compound-containing medium was replaced daily for up to 7 days. Electrophysiological properties of the employed cell types were recorded by automated patch-clamp (Patchliner) and the results were integrated into the electromechanical model of the system.Calcium channel agonist S Bay K8644 and beta-adrenergic stimulator isoproterenol induced significant positive inotropic responses without additional external stimulation. Kinase inhibitors displayed cardiotoxic effects on a functional level at low concentrations. The system-integrated analysis detected alterations in beating shape as well as frequency and arrhythmic events and we provide a quantitative measure of these.
Y1  - 2020
UR  - https://opus.bibliothek.fh-aachen.de/opus4/frontdoor/index/index/docId/9201
SN  - 0006-3495
N1  - Raman, Arayind Hariharan 
im Artikel unter dem Namen: Raman, Alexander H.
VL  - 118
IS  - Issue 3, Supplement 1
SP  - 570a
PB  - Elsevier
CY  - Amsterdam
ER  -