M Marziano1, S Tonello2, E Cantù2, G Abate3, M Vezzoli3, W Rungratanawanich3, M Serpelloni2, N F Lopomo2, M Memo3, E Sardini2, D Uberti3. 1. Department of Information Engineering, University of Brescia, Brescia, Italy; Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy. Electronic address: m.marziano@unibs.it. 2. Department of Information Engineering, University of Brescia, Brescia, Italy. 3. Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy.
Abstract
BACKGROUND: Colorectal adenocarcinoma cells (Caco-2) are a widely used model of intestinal barrier to study cancer development, toxicological assessments, absorption and metabolism in food science or drug discovery. Caco-2 spontaneously differentiate into a monolayer expressing several specific characteristics, typically showed by mature enterocytes. For in vitro experiments, it is crucial to identify non-invasive and non-destructive techniques able to evaluate the integrity and differentiation of the cells monolayer. Thus, we aimed to assess these properties by analyzing electrical impedance measurements. METHODS: Caco-2 cells were differentiated for 21 days. The monolayer integrity and differentiation were primarily evaluated by means of morphological, biochemical and molecular data. Impedance measurements in a range of frequencies from 400 Hz to 50 kHz were performed using a dedicated set up, including customized Aerosol Jet Printed carbon-based sensors. RESULTS: The trends of RI observed at three different frequencies were able to describe cell growth and differentiation. In order to evaluate which frequencies better correlate with cell differentiation, Principal Component Analysis have been employed and the concordance analysis between RI magnitude and morphological, biochemical and molecular data, highlighted 40 kHz as the optimal frequency to assess Caco-2 cells differentiation process. CONCLUSION: We demonstrated the feasibility and reliability of applying impedance-based measurements not only to provide information about the monolayer status, but also for cell differentiation monitoring. GENERAL SIGNIFICANCE: This study underlined the possibility to use a dedicated sensor to assess the integrity and differentiation of Caco-2 monolayer, as a reliable non-destructive alternative to conventional approaches.
BACKGROUND:Colorectal adenocarcinoma cells (Caco-2) are a widely used model of intestinal barrier to study cancer development, toxicological assessments, absorption and metabolism in food science or drug discovery. Caco-2 spontaneously differentiate into a monolayer expressing several specific characteristics, typically showed by mature enterocytes. For in vitro experiments, it is crucial to identify non-invasive and non-destructive techniques able to evaluate the integrity and differentiation of the cells monolayer. Thus, we aimed to assess these properties by analyzing electrical impedance measurements. METHODS:Caco-2 cells were differentiated for 21 days. The monolayer integrity and differentiation were primarily evaluated by means of morphological, biochemical and molecular data. Impedance measurements in a range of frequencies from 400 Hz to 50 kHz were performed using a dedicated set up, including customized Aerosol Jet Printed carbon-based sensors. RESULTS: The trends of RI observed at three different frequencies were able to describe cell growth and differentiation. In order to evaluate which frequencies better correlate with cell differentiation, Principal Component Analysis have been employed and the concordance analysis between RI magnitude and morphological, biochemical and molecular data, highlighted 40 kHz as the optimal frequency to assess Caco-2 cells differentiation process. CONCLUSION: We demonstrated the feasibility and reliability of applying impedance-based measurements not only to provide information about the monolayer status, but also for cell differentiation monitoring. GENERAL SIGNIFICANCE: This study underlined the possibility to use a dedicated sensor to assess the integrity and differentiation of Caco-2 monolayer, as a reliable non-destructive alternative to conventional approaches.
Authors: Min Seo Jeon; Yoon Young Choi; Sung Jun Mo; Jang Ho Ha; Young Seo Lee; Hee Uk Lee; Soo Dong Park; Jae-Jung Shim; Jung-Lyoul Lee; Bong Geun Chung Journal: Nano Converg Date: 2022-02-08
Authors: Dahiana Mojena-Medina; Moritz Hubl; Manuel Bäuscher; José Luis Jorcano; Ha-Duong Ngo; Pablo Acedo Journal: Sensors (Basel) Date: 2020-10-08 Impact factor: 3.576