Development of a multiparametric (bio)sensing platform for continuous monitoring of stress metabolites

Abstract

There is a growing need for real-time monitoring of metabolic products that could reflect cell damages over extended periods. In this paper, we report the design and development of an original multiparametric (bio)sensing platform that is tailored for the real-time monitoring of cell metabolites derived from cell cultures. Most attractive features of our developed electrochemical (bio)sensing platform are its easy manufacturing process, that enables seamless scale-up, modular and versatile approach, and low cost. In addition, the developed platform allows a multiparametric analysis instead of single-analyte analysis. Here we provide an overview of the sensors-based analysis of four main factors that can indicate a possible cell deterioration problem during cell-culture: pH, hydrogen peroxide, nitric oxide/nitrite and lactate. Herein, we are proposing a sensors platform based on thick-film coupled to microfluidic technology that can be integrated into any microfluidic system using Luer-lock connectors. This platform allows obtaining an accurate analysis of the secreting stress metabolites during cell/tissues culture.

Characterization of the Impact of Classical Cell‐culture Media on the Response of Electrochemical Sensors

“…This may explain the higher sensitivity to H 2 O 2 in William's medium [27]. So far, comparing to H 2 O 2 detection in PBS medium, the hydrogen peroxide sensors show a close detection limit in cell culture media under hydrodynamic conditions and are estimated to be a few μM as already reported in our previous work (in the range of 8–20 μM) [4].…”

“…Lactate levels were recorded using amperometric enzymatic biosensors (electrodes 5 and 6) that embed the enzyme lactate oxidase (LOx) [4, 32]. LOx is immobilized at the surface of a PB electrode through crosslinking using Glutaraldehyde and BSA.…”

“…The 2D and 3D design of the multiparametric electrochemical (bio)sensing platform has already been described in our previous work [4]. Briefly, this printed circuit board (PCB) integrates one reference electrode (RE, 3 mm), one annular counter‐electrode (CE, Internal diameter of 8.46 mm, External Diameter of 9.85 mm) and eight working electrodes (WE, diameter of 1.6 mm) (Figure 1a).…”

“…For the multiparametric (bio)sensing platform, four parameters are monitored as follows: pH, H 2 O 2 , NO/NO 2 − and lactate. For these parameters, the working electrodes are functionalized by pairs by screen‐printing or drop casting of the different sensitive layers under clean room conditions [4]. In terms of the electrodes materials, our strategy consists of selecting processable materials in the view of industrializing the (bio)sensing platform.…”

“…More particularly, the PANBioRA system integrates a cytotoxicity-assessment microfluidic-module composed of a cell culture chamber connected to two analytical modules enabling respectively the monitoring of inflammatory biomarkers and of metabolites related to the cell stress [1][2][3]. The latter (including NO, lactate, pH and H 2 O 2 ) are monitored in line using a multiparametric electroanalytical platform [4]. However, according to the cultured cells, specific media that differs widely by their composition have to be used [5] and might significantly impact the electrochemical (bio)sensors response.…”