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Muscle-on-a-chip with an on-site multiplexed biosensing system for in situ monitoring of secreted IL-6 and TNF-α.

Lab on a chip (2019-06-28)
María A Ortega, Xiomara Fernández-Garibay, Albert G Castaño, Francesco De Chiara, Alejandro Hernández-Albors, Jordina Balaguer-Trias, Javier Ramón-Azcón
ABSTRACT

Despite the increasing number of organs-on-a-chip that have been developed in the past decade, limited efforts have been made to integrate a sensing system for in situ continual measurements of biomarkers from three-dimensional (3D) tissues. Here, we present a custom-made integrated platform for muscle cell stimulation under fluidic conditions connected with a multiplexed high-sensitivity electrochemical sensing system for in situ monitoring. To demonstrate this, we use our system to measure the release levels and release time of interleukin 6 and tumor necrosis factor alpha in vitro by 3D muscle microtissue under electrical and biological stimulations. Our experimental design has enabled us to perform multiple time point measurements using functionalized screen-printed gold electrodes with sensitivity in the ng mL-1 range. This affordable setup is uniquely suited for monitoring factors released by 3D single cell types upon external stimulation for metabolic studies.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
N-Hydroxysuccinimide, 98%
Sigma-Aldrich
MES hydrate, ≥99.5% (titration), pH 2.5-4.0 (0.5 M in H2O), BioXtra
Sigma-Aldrich
Poly(dimethylsiloxane), viscosity 1.0 cSt (25 °C)
Sigma-Aldrich
Formalin solution, neutral buffered, 10%, histological tissue fixative
Sigma-Aldrich
Sodium carboxymethyl cellulose, average Mw ~90,000
Sigma-Aldrich
N-Hydroxysulfosuccinimide sodium salt, ≥98% (HPLC)