A Bioreactor to Apply Multimodal Physical Stimuli to Cultured Cells

Abstract

SummaryCells residing in the cardiac niche are constantly experiencing physical stimuli, including electrical pulses and cyclic mechanical stretch. These physical signals are known to influence a variety of cellular functions, including the secretion of growth factors and extracellular matrix proteins in cardiac fibroblasts, calcium handling and contractility in cardiomyocytes, or stretch-activated ion channels in muscle and non-muscle cells of the cardiovascular system. Recent progress in cardiac tissue engineering suggests that controlled physical stimulation can lead to functional improvements in multicellular cardiac tissue constructs. To study these effects, aspects of the physical environment of the myocardium have to be mimicked in vitro. This protocol demonstrates how a specifically designed bioreactor system allows controlled exposure of cultured cells, during continuous live imaging, to cyclic stretch, rhythmic electrical stimulation, fluid perfusion, at user-defined temperatures.

Consecutive-Day Ventricular and Atrial Cardiomyocyte Isolations from the Same Heart: Shifting the Cost–Benefit Balance of Cardiac Primary Cell Research

“…In addition, a host of supplements have been previously suggested to enhance the survival of cultured CM—thyroid hormones, insulin, L-glutamate, creatine, L-carnitine, and taurine [ 21 , 31 ]. Finally, the viability of slices could be extended by culturing under constant electro-mechanical stimulation (shown to preserve slice function) between isolations [ 12 , 32 , 33 , 34 ].…”

Electrophysiological Stimulation of Whole Heart Constructs in an 8‐Pole Electrical Field

“…In the most recent past a substantial amount of published work on electrophysiological stimulation of myocardial constructs has demonstrated the feasibility of the here applied concept, thereby providing well founded information for scientific applications and their technical realization . The herein presented principle of an eight‐pole field is a novel development that delivers new options for complex three‐dimensional stimulation of WHC.…”

“…Here we show a concept for electrical field stimulation for the whole heart approach as an electrical stimulation principle alternative to direct current injection, with an application to WHC installed within the previously elaborated bioreactor system. Today, electrical field stimulation is a proven and established concept in myocardial tissue engineering and has been applied to various combinations of cell‐scaffold constructions . While electrodes that are contacted or plugged into densely reseeded constructs enable a direct and straight electrical current flow, provoking a robust overall polarization, we believe that a multipolar electrical field may conform the complex micro architecture of the whole heart scaffolds more closely.…”