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Electric Cell-Substrate Impedance Sensing (ECIS) with Microelectrode Arrays for Investigation of Cancer Cell-Fibroblasts Interaction.

Tran TB, Baek C, Min J - PLoS ONE (2016)

Bottom Line: In brief, a co-culture device consisting of 2 individual fluidic chambers in parallel, which were separated by a 100 μm fence was utilized for cell patterning.Microelectrodes arrays were installed within each chamber including electrodes at various distances away from the confrontation line for the electrochemical impedimetric sensing assessment of cell-to-cell influence.After the fence was removed and cell-to-cell contact occurred, by evaluating the impedance signal responses representing cell condition and behavior, both direct and indirect cell-to-cell interactions through conditioned media were investigated.

View Article: PubMed Central - PubMed

Affiliation: School of Integrative Engineering, Chung-Ang University, Heukseok-dong, Dongjak-gu, Seoul, Republic of Korea.

ABSTRACT
The tumor microenvironment, including stromal cells, surrounding blood vessels and extracellular matrix components, has been defined as a crucial factor that influences the proliferation, drug-resistance, invasion and metastasis of malignant epithelial cells. Among other factors, the communications and interaction between cancer cells and stromal cells have been reported to play pivotal roles in cancer promotion and progression. To investigate these relationships, an on-chip co-culture model was developed to study the cellular interaction between A549-human lung carcinoma cells and MRC-5-human lung epithelial cells in both normal proliferation and treatment conditions. In brief, a co-culture device consisting of 2 individual fluidic chambers in parallel, which were separated by a 100 μm fence was utilized for cell patterning. Microelectrodes arrays were installed within each chamber including electrodes at various distances away from the confrontation line for the electrochemical impedimetric sensing assessment of cell-to-cell influence. After the fence was removed and cell-to-cell contact occurred, by evaluating the impedance signal responses representing cell condition and behavior, both direct and indirect cell-to-cell interactions through conditioned media were investigated. The impact of specific distances that lead to different influences of fibroblast cells on cancer cells in the co-culture environment was also defined.

No MeSH data available.


Related in: MedlinePlus

(A) Microelectrode arrays (1: working electrode, 2: common counter electrode) were fabricated on experimental glass slides (76mm x 52mm) by common photolithography processes. SU-8 photoresist was used as passivation layer to cover the conducting traces. (B) The sensing platform was divided into 2 areas, one for the cancer cells and one for microenvironment agents. Several mirco-sized working electrodes were installed in each area at different distances: 100, 250, 650, 1450 and 3050μm far from the confrontation line. (C) A single electrode was 100μm in diameter. (D) An image of a real chip after the co-culturing of 2 different cell types on 2 sides. (E) The co-culture patterning process using a dual-chamber mold. (E1) After treating the surface for cell culture, the cell chip was placed on the designed fixture, and the dual-chamber mold was fixed at the proper location by screws. The 2 chambers were separated each other by a 100μm thick wall for cell patterning. (E2) After the cells in both chamber had attached to the chip surface, the dual-chamber mold was replaced by a well-type open reservoir. A PDMS bed was also installed to prevent solution from leaking. The cell chip was connected to the measurement system and was kept in the incubator during the measurements.
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pone.0153813.g001: (A) Microelectrode arrays (1: working electrode, 2: common counter electrode) were fabricated on experimental glass slides (76mm x 52mm) by common photolithography processes. SU-8 photoresist was used as passivation layer to cover the conducting traces. (B) The sensing platform was divided into 2 areas, one for the cancer cells and one for microenvironment agents. Several mirco-sized working electrodes were installed in each area at different distances: 100, 250, 650, 1450 and 3050μm far from the confrontation line. (C) A single electrode was 100μm in diameter. (D) An image of a real chip after the co-culturing of 2 different cell types on 2 sides. (E) The co-culture patterning process using a dual-chamber mold. (E1) After treating the surface for cell culture, the cell chip was placed on the designed fixture, and the dual-chamber mold was fixed at the proper location by screws. The 2 chambers were separated each other by a 100μm thick wall for cell patterning. (E2) After the cells in both chamber had attached to the chip surface, the dual-chamber mold was replaced by a well-type open reservoir. A PDMS bed was also installed to prevent solution from leaking. The cell chip was connected to the measurement system and was kept in the incubator during the measurements.

Mentions: In this study, we proposed a simple co-culture model with embedded high-throughput microelectrode arrays (MEA) using an electric cell-substrate impedance sensing (ECIS) assay (Fig 1) to monitor tumor cell conditions continuously when confronted with cultured fibroblasts. This electrical sensing method was utilized in this study due to the prominent advantages; this method is non-invasive, simple to setup, easy to perform, extraordinarily sensitive to cellular conditions and capable of real-time monitoring [16,17]. The MEA were patterned on both sides of the cell culture areas at various distances from the separation line. During the cell interactions, each electrode continuously records an impedance signal through a high-throughput data acquisition system. This type of impedimetric data has been acknowledged to reflect cellular adhesion, spreading, proliferation and viability in treatment conditions with environmental toxins, drugs, and chemicals, as well as other substances.


Electric Cell-Substrate Impedance Sensing (ECIS) with Microelectrode Arrays for Investigation of Cancer Cell-Fibroblasts Interaction.

Tran TB, Baek C, Min J - PLoS ONE (2016)

(A) Microelectrode arrays (1: working electrode, 2: common counter electrode) were fabricated on experimental glass slides (76mm x 52mm) by common photolithography processes. SU-8 photoresist was used as passivation layer to cover the conducting traces. (B) The sensing platform was divided into 2 areas, one for the cancer cells and one for microenvironment agents. Several mirco-sized working electrodes were installed in each area at different distances: 100, 250, 650, 1450 and 3050μm far from the confrontation line. (C) A single electrode was 100μm in diameter. (D) An image of a real chip after the co-culturing of 2 different cell types on 2 sides. (E) The co-culture patterning process using a dual-chamber mold. (E1) After treating the surface for cell culture, the cell chip was placed on the designed fixture, and the dual-chamber mold was fixed at the proper location by screws. The 2 chambers were separated each other by a 100μm thick wall for cell patterning. (E2) After the cells in both chamber had attached to the chip surface, the dual-chamber mold was replaced by a well-type open reservoir. A PDMS bed was also installed to prevent solution from leaking. The cell chip was connected to the measurement system and was kept in the incubator during the measurements.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835071&req=5

pone.0153813.g001: (A) Microelectrode arrays (1: working electrode, 2: common counter electrode) were fabricated on experimental glass slides (76mm x 52mm) by common photolithography processes. SU-8 photoresist was used as passivation layer to cover the conducting traces. (B) The sensing platform was divided into 2 areas, one for the cancer cells and one for microenvironment agents. Several mirco-sized working electrodes were installed in each area at different distances: 100, 250, 650, 1450 and 3050μm far from the confrontation line. (C) A single electrode was 100μm in diameter. (D) An image of a real chip after the co-culturing of 2 different cell types on 2 sides. (E) The co-culture patterning process using a dual-chamber mold. (E1) After treating the surface for cell culture, the cell chip was placed on the designed fixture, and the dual-chamber mold was fixed at the proper location by screws. The 2 chambers were separated each other by a 100μm thick wall for cell patterning. (E2) After the cells in both chamber had attached to the chip surface, the dual-chamber mold was replaced by a well-type open reservoir. A PDMS bed was also installed to prevent solution from leaking. The cell chip was connected to the measurement system and was kept in the incubator during the measurements.
Mentions: In this study, we proposed a simple co-culture model with embedded high-throughput microelectrode arrays (MEA) using an electric cell-substrate impedance sensing (ECIS) assay (Fig 1) to monitor tumor cell conditions continuously when confronted with cultured fibroblasts. This electrical sensing method was utilized in this study due to the prominent advantages; this method is non-invasive, simple to setup, easy to perform, extraordinarily sensitive to cellular conditions and capable of real-time monitoring [16,17]. The MEA were patterned on both sides of the cell culture areas at various distances from the separation line. During the cell interactions, each electrode continuously records an impedance signal through a high-throughput data acquisition system. This type of impedimetric data has been acknowledged to reflect cellular adhesion, spreading, proliferation and viability in treatment conditions with environmental toxins, drugs, and chemicals, as well as other substances.

Bottom Line: In brief, a co-culture device consisting of 2 individual fluidic chambers in parallel, which were separated by a 100 μm fence was utilized for cell patterning.Microelectrodes arrays were installed within each chamber including electrodes at various distances away from the confrontation line for the electrochemical impedimetric sensing assessment of cell-to-cell influence.After the fence was removed and cell-to-cell contact occurred, by evaluating the impedance signal responses representing cell condition and behavior, both direct and indirect cell-to-cell interactions through conditioned media were investigated.

View Article: PubMed Central - PubMed

Affiliation: School of Integrative Engineering, Chung-Ang University, Heukseok-dong, Dongjak-gu, Seoul, Republic of Korea.

ABSTRACT
The tumor microenvironment, including stromal cells, surrounding blood vessels and extracellular matrix components, has been defined as a crucial factor that influences the proliferation, drug-resistance, invasion and metastasis of malignant epithelial cells. Among other factors, the communications and interaction between cancer cells and stromal cells have been reported to play pivotal roles in cancer promotion and progression. To investigate these relationships, an on-chip co-culture model was developed to study the cellular interaction between A549-human lung carcinoma cells and MRC-5-human lung epithelial cells in both normal proliferation and treatment conditions. In brief, a co-culture device consisting of 2 individual fluidic chambers in parallel, which were separated by a 100 μm fence was utilized for cell patterning. Microelectrodes arrays were installed within each chamber including electrodes at various distances away from the confrontation line for the electrochemical impedimetric sensing assessment of cell-to-cell influence. After the fence was removed and cell-to-cell contact occurred, by evaluating the impedance signal responses representing cell condition and behavior, both direct and indirect cell-to-cell interactions through conditioned media were investigated. The impact of specific distances that lead to different influences of fibroblast cells on cancer cells in the co-culture environment was also defined.

No MeSH data available.


Related in: MedlinePlus