Doxorubicin‐induced cardiomyocyte toxicity – protective effects of endothelial cells in a tri‐culture model system

Abstract

Doxorubicin-induced cardiomyopathy is a clinically prevalent pathology, occurring as a sequelae following chemotherapy for cancer patients. In particular, the "first dose" effect has been particularly challenging, given the heterogeneous and multifactorial nature of this pathophysiology. Here, we describe the development of a physiologically relevant in vitro model for cardiotoxicity testing, using human cells. Primary cardiomyocytes, endothelial, and smooth muscle cells were tri-cultured in 2D, or within nano-fibrous scaffolds in a 3D environment, under dynamic nutrient flow, using the Quasi Vivo® system. State-ofthe-art sensor chips were used to detect troponin I levels, 2 h after acute exposure to doxorubicin. We demonstrate a significant improvement in cardiomyocyte viability when grown in a 3D tri-culture environment over a 5-day period and a 10-fold reduction in doxorubicin-induced toxicity. Our tri-culture model can be used as a valuable tool for physiologically relevant assessment of drug-induced cardiotoxicity in vitro.

Differential effects of resveratrol on the dilator responses of femoral arteries, ex vivo

“…Cells were seeded onto Nunc Thermanox coverslips (10,000 cells/coverslip; 13 mm diameter) (Thermo Fisher Scientific, USA) and allowed to proliferate for 24 hours (approximately 90% confluency). Half of the coverslips were kept in the well plate to perform the ACh assay while the other overslips were transferred into a millifluidic system made up of a bioreactor chamber (Kirkstall, UK) attached with a flow pump system in which increased flow was introduced [41] as indicated in the section below. All reagents were obtained from Sigma-Aldrich (Poole, UK), unless otherwise stated.…”

“…Control cultures were treated with growth medium (0.1% DMSO) alone. After the incubation, the supernatant was collected and the cultured cells exposed to ACh 10 -5 M for 2-10 minutes, or flow rates of 5-10 Lmin -1 , calculated at 5.59 x 10 -4 dyn/cm 2 (representing physiological rates of interstitial flow) [41], for 3-9 minutes. Following exposure to ACh or increased flow, the supernatant was collected and the production of NO quantified using the Griess assay (23479, Sigma Aldrich, UK).…”

Biomimetic microsystems for cardiovascular studies

“…The plug and play nature of the system offers broad flexibility and reduces some of the complications associated with linking multiple-chip models. Quasi Vivo is a commercially available platform that offers the versatility to connect different cell chambers in various combinations (215). Human cardiomyocytes, endothelial cells, and smooth muscle cells were cultured in a 3-D poly-vinyl alcohol-gelatin nanofibrous scaffold in the Quasi Vivo system to model doxorubicin-induced cardiotoxicity.…”