Presentation Information
[P01-104]Quantifying Glucose, Lactic Acid, and Chloride Ion Dynamics to Evaluate Drug Performance in In Vitro Intestinal Models
○RUITONG ZHU1, Yoshinobu Utagawa1, Hiroya Abe1,2, Hitoshi Shiku1, Kosuke Ino1 (1. Grad. Sch. of Eng., Tohoku Univ. (Japan), 2. FRIS, Tohoku Univ. (Japan))
Keywords:
Microphysiological System,Transwell Chamber,Intestinal Model,Drug Evaluation,Biosensing
[Purpose]
Microphysiological systems (MPS) are essential for mimicking human in vivo functions, particularly in intestinal research. This study aims to construct robust intestinal models and perform comprehensive quantitative analyses of cell-derived chemicals. Compared to traditional transepithelial electrical resistance (TEER) for physical barrier monitoring, we incorporate quantitative analyses of glucose, lactic acid, and chloride ions. These markers enable the evaluation of metabolic activity and ion transport functions, providing a more comprehensive physiological framework for assessing drug behavior.
[Method]
To construct and analyze intestinal models, Caco-2 cells were cultured on porous membranes in Transwell chambers with daily medium changes to ensure stability. Following the culture period, electrochemical methods were used to quantify metabolites and ions, specifically glucose, lactic acid, Cl-, Na+, and K+, collected from both apical and basolateral compartments.Drug responses were evaluated using Afatinib and forskolin, with CFTR(inh)-172 utilized as a specific forskolin inhibitor. Furthermore, transmembrane ion transport was assessed by applying a Cl- concentration gradient across the cell layer.
[Results]
Caco-2 cell layers exhibited consistent metabolic capacity after four days, achieving complete glucose consumption within 24 hours. While Afatinib and forskolin did not cause immediate cytotoxicity or cell detachment, they significantly altered glucose consumption and lactic acid production rates. Conversely, Cl- permeability showed only minor changes upon drug exposure. These findings demonstrate that cellular metabolic activity is more sensitive to drug exposure than TEER.
[Discussion]
The distinct sensitivity profiles demonstrate that metabolic profiling provides a more robust framework for characterizing drug-induced cellular responses. Because glucose consumption and lactic acid production respond more dynamically to pharmaceutical compounds than Cl- permeability, these metabolic outputs function as more effective, early-warning biomarkers. This approach moves beyond permeability assays to evaluate functional metabolic health, allowing for the detection of subtle drug effects even when barrier integrity remains intact.
[Conclusion]
This study presents a comprehensive analytical framework for evaluating engineered intestinal models using electrochemical methods. To further enhance physiological relevance, future research will focus on developing co-culture microfluidic devices and integrating electrochemical sensors for real-time, in situ monitoring of the cellular microenvironment.
Microphysiological systems (MPS) are essential for mimicking human in vivo functions, particularly in intestinal research. This study aims to construct robust intestinal models and perform comprehensive quantitative analyses of cell-derived chemicals. Compared to traditional transepithelial electrical resistance (TEER) for physical barrier monitoring, we incorporate quantitative analyses of glucose, lactic acid, and chloride ions. These markers enable the evaluation of metabolic activity and ion transport functions, providing a more comprehensive physiological framework for assessing drug behavior.
[Method]
To construct and analyze intestinal models, Caco-2 cells were cultured on porous membranes in Transwell chambers with daily medium changes to ensure stability. Following the culture period, electrochemical methods were used to quantify metabolites and ions, specifically glucose, lactic acid, Cl-, Na+, and K+, collected from both apical and basolateral compartments.Drug responses were evaluated using Afatinib and forskolin, with CFTR(inh)-172 utilized as a specific forskolin inhibitor. Furthermore, transmembrane ion transport was assessed by applying a Cl- concentration gradient across the cell layer.
[Results]
Caco-2 cell layers exhibited consistent metabolic capacity after four days, achieving complete glucose consumption within 24 hours. While Afatinib and forskolin did not cause immediate cytotoxicity or cell detachment, they significantly altered glucose consumption and lactic acid production rates. Conversely, Cl- permeability showed only minor changes upon drug exposure. These findings demonstrate that cellular metabolic activity is more sensitive to drug exposure than TEER.
[Discussion]
The distinct sensitivity profiles demonstrate that metabolic profiling provides a more robust framework for characterizing drug-induced cellular responses. Because glucose consumption and lactic acid production respond more dynamically to pharmaceutical compounds than Cl- permeability, these metabolic outputs function as more effective, early-warning biomarkers. This approach moves beyond permeability assays to evaluate functional metabolic health, allowing for the detection of subtle drug effects even when barrier integrity remains intact.
[Conclusion]
This study presents a comprehensive analytical framework for evaluating engineered intestinal models using electrochemical methods. To further enhance physiological relevance, future research will focus on developing co-culture microfluidic devices and integrating electrochemical sensors for real-time, in situ monitoring of the cellular microenvironment.
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