Real-time monitoring of relaxation and contractility of smooth muscle cells on a novel biohybrid chip

Abstract

Cardiovascular diseases represent the most common cause of death in industrialized countries. In this context vascular smooth muscle cells (SMCs) are a major key player that is involved in pathological processes like hypertension and atherosclerosis. Therefore the pharmaceutical industry is intensively investigated in developing non-destructive and label-free monitoring techniques for a quantitative detection of SMC characteristics in the field of active pharmaceutical development as well as clinical diagnostics. Hence, we developed a novel multiwell interdigital electrode sensor-array in standardized ANSI 96-well layout. Through optimization of electrode geometry and material as well as passivation/adhesion-layer we obtained a novel biohybrid chip for the sensitive and quantitative detection of SMC contractility as well as relaxation via impedance spectroscopy. For the validation of our multiwell sensor-array we established a SMC culture model derived from primary cells that is switchable from a non-contractile pathological to a functional contractile phenotype. Using the reference compounds acetylcholine (ACh) and amlodipine, we could quantify SMC contraction by an impedance decrease to 40% while SMC relaxation was detectable by an impedance increase to 110%. More strikingly we could monitor aging of the isolated SMC which arose by an attenuated contractility over successive passaging. Demonstrating the performance of our self-developed multiwell sensor-array based impedance measurement setup we provide a suitable sensor-array coupled cell model to study the mechanisms that activated SMCs undergo in response to inflammatory mediators or vessel injury.

A novel 96-well multielectrode array based impedimetric monitoring platform for comparative drug efficacy analysis on 2D and 3D brain tumor cultures

“…In this way, up to 800 spheroids can be measured by an individual experimenter on a single chip and measurement system per day (Seidel et al, 2012). In contrast, small scale multiwell-electrode arrays (Haas et al, 2010) with adhered 2D cultures are automatically and continuously measured with a single measurement setup, throughout the whole experiment. To overcome the limitation of a small number of wells in these arrays, we upscaled the planar array to an ANSI compatible 96-well array (Fig.…”

“…To perform a screening on several tumor cell lines as model of patient specific tumors for determination of pharmacokinetic relevant parameters, the throughput had to be increased. Therefore, we upscaled the planar multielectrode arrays (Haas et al, 2010;Poenick et al, 2014) to an ANSI compatible full 96-well-multielectrode array. In combination with our well-established microcavity arrays for the monitoring of 3D cultures, a comprehensive impedimetric screening platform could be established.…”

Quantitative impedimetric NPY-receptor activation monitoring and signal pathway profiling in living cells

“…Interdigital electrode arrays comprising nine wells in a 96-well format were produced by lift of technique as previously described (Haas et al, 2010). For a combined impedimetric and optical monitoring of adherent cell layers the electrode finger geometry was optimized (electrode finger width 200 mm, distance between electrode fingers 250 mm).…”

“…In the present study, we investigated the possibility for direct NPY-receptor activation monitoring by impedance spectroscopy using an optimized interdigital electrode array based on an initial prototype (Haas et al, 2010) and which molecular alterations are responsible for the impedance change.…”

Direct Chemosensitivity Monitoring Ex Vivo on Undissociated Melanoma Tumor Tissue by Impedance Spectroscopy

“…Impedimetric measurements on 2D cultures were performed as previously described (20). Briefly, 5 Â 10 4 melanoma cells were seeded in 200-mL complete growth medium per well on self-developed interdigital electrode (IDE) arrays coated with collagen I (Life Technologies) and cultivated for 3 to 4 days until cell layer confluence.…”