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Study Hypoxic Response under Cyclic Oxygen Gradients Generated in Microfluidic Devices Using Real-Time Fluorescence Imaging

Oxygen plays important roles in regulating various biological activities under physiological and pathological conditions. However, the response of cells facing temporal variation in oxygen microenvironments has seldom been studied due to technical limitations. In this paper, an integrated approach t...

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Detalles Bibliográficos
Autores principales: Chang, Dao-Ming, Tung, Yi-Chung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9688408/
https://www.ncbi.nlm.nih.gov/pubmed/36421149
http://dx.doi.org/10.3390/bios12111031
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author Chang, Dao-Ming
Tung, Yi-Chung
author_facet Chang, Dao-Ming
Tung, Yi-Chung
author_sort Chang, Dao-Ming
collection PubMed
description Oxygen plays important roles in regulating various biological activities under physiological and pathological conditions. However, the response of cells facing temporal variation in oxygen microenvironments has seldom been studied due to technical limitations. In this paper, an integrated approach to studying hypoxic response under cyclic oxygen gradients is developed. In the experiments, a cell culture system based on a microfluidic device is constructed to generate cyclic oxygen gradients with desired periods by alternately introducing gases with specific compositions into the microfluidic channels next to the cell culture channel separated by thin channel walls. Observation of the hypoxic responses is performed using real-time fluorescence imaging of dyes sensitive to extra- and intracellular oxygen tensions as well as intracellular calcium concentrations. Cellular hypoxic responses of human aortic smooth muscle cells (AoSMCs) and lung carcinoma epithelium (A549) cells, including intracellular oxygen and calcium levels, are measured. The results show that the two types of cells have different hypoxic responses to the applied cyclic oxygen gradients. With the capability of real-time cellular response monitoring under cyclic oxygen gradients, the developed approach provides a useful scheme to investigate hypoxic responses in vitro under microenvironments mimicking various in vivo physiological and pathological conditions.
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spelling pubmed-96884082022-11-25 Study Hypoxic Response under Cyclic Oxygen Gradients Generated in Microfluidic Devices Using Real-Time Fluorescence Imaging Chang, Dao-Ming Tung, Yi-Chung Biosensors (Basel) Article Oxygen plays important roles in regulating various biological activities under physiological and pathological conditions. However, the response of cells facing temporal variation in oxygen microenvironments has seldom been studied due to technical limitations. In this paper, an integrated approach to studying hypoxic response under cyclic oxygen gradients is developed. In the experiments, a cell culture system based on a microfluidic device is constructed to generate cyclic oxygen gradients with desired periods by alternately introducing gases with specific compositions into the microfluidic channels next to the cell culture channel separated by thin channel walls. Observation of the hypoxic responses is performed using real-time fluorescence imaging of dyes sensitive to extra- and intracellular oxygen tensions as well as intracellular calcium concentrations. Cellular hypoxic responses of human aortic smooth muscle cells (AoSMCs) and lung carcinoma epithelium (A549) cells, including intracellular oxygen and calcium levels, are measured. The results show that the two types of cells have different hypoxic responses to the applied cyclic oxygen gradients. With the capability of real-time cellular response monitoring under cyclic oxygen gradients, the developed approach provides a useful scheme to investigate hypoxic responses in vitro under microenvironments mimicking various in vivo physiological and pathological conditions. MDPI 2022-11-17 /pmc/articles/PMC9688408/ /pubmed/36421149 http://dx.doi.org/10.3390/bios12111031 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Chang, Dao-Ming
Tung, Yi-Chung
Study Hypoxic Response under Cyclic Oxygen Gradients Generated in Microfluidic Devices Using Real-Time Fluorescence Imaging
title Study Hypoxic Response under Cyclic Oxygen Gradients Generated in Microfluidic Devices Using Real-Time Fluorescence Imaging
title_full Study Hypoxic Response under Cyclic Oxygen Gradients Generated in Microfluidic Devices Using Real-Time Fluorescence Imaging
title_fullStr Study Hypoxic Response under Cyclic Oxygen Gradients Generated in Microfluidic Devices Using Real-Time Fluorescence Imaging
title_full_unstemmed Study Hypoxic Response under Cyclic Oxygen Gradients Generated in Microfluidic Devices Using Real-Time Fluorescence Imaging
title_short Study Hypoxic Response under Cyclic Oxygen Gradients Generated in Microfluidic Devices Using Real-Time Fluorescence Imaging
title_sort study hypoxic response under cyclic oxygen gradients generated in microfluidic devices using real-time fluorescence imaging
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9688408/
https://www.ncbi.nlm.nih.gov/pubmed/36421149
http://dx.doi.org/10.3390/bios12111031
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