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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) Resistance responses of MEA to various concentrations of A549 tumor cells in monoculture for 16 h (at 10mV, 10kHz). (B) This calibration curve represents the correlation between A549 cell density and cell index value. (C) MTS results from of monocultures of A549 cells and MRC-5 fibroblasts after 72 h of exposure to curcumin. The IC50 values for the A549 cells and MRC-5 cells were 50 μM and 30 μM, respectively. The data are shown as the mean (±SD) of 3 repetitions.
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pone.0153813.g002: (A) Resistance responses of MEA to various concentrations of A549 tumor cells in monoculture for 16 h (at 10mV, 10kHz). (B) This calibration curve represents the correlation between A549 cell density and cell index value. (C) MTS results from of monocultures of A549 cells and MRC-5 fibroblasts after 72 h of exposure to curcumin. The IC50 values for the A549 cells and MRC-5 cells were 50 μM and 30 μM, respectively. The data are shown as the mean (±SD) of 3 repetitions.

Mentions: In the first experimental step, the MEA cell chips were validated via monocultures of A549 tumor cells and MRC-5 fibroblasts in normal proliferative conditions. Before operating the cell measurement system, the arrays were adjusted (by LabVIEW) to normalize the systematic differences, such as wiring, soldering and conductive traces. The normalized cell-free resistance of each electrode was fixed at 2.6 kΩ. Fig 2A shows the resistance responses when single electrodes were challenged by various concentrations of A549 cells. The data obtained over 16 h were fitted by a hill algorithm for a growth/sigmoidal model (OriginPro 9). Below 1.25×105 cells/well, the resistance curves corresponded well to the fitting curves. At higher cell densities, a lag phase appeared from 4 h to 8 h, when the cells were rearranging themselves on binding the site prior to forming a monolayer on the electrodes. To easily estimate viability throughout this study, a dimensionless parameter termed the cell index (CI) was defined as the relative change in measured electrical resistance and can be calculated using the equation belowCI=RiRcell−free−1where Ri is the resistance of the cell-covered electrodes, and Rcell-free is the resistance of blank electrodes (2.6 kΩ). After 16 h, a calibration curve was plotted, which represented the correlation between CI value and cell seeding density (Fig 2B).


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) Resistance responses of MEA to various concentrations of A549 tumor cells in monoculture for 16 h (at 10mV, 10kHz). (B) This calibration curve represents the correlation between A549 cell density and cell index value. (C) MTS results from of monocultures of A549 cells and MRC-5 fibroblasts after 72 h of exposure to curcumin. The IC50 values for the A549 cells and MRC-5 cells were 50 μM and 30 μM, respectively. The data are shown as the mean (±SD) of 3 repetitions.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153813.g002: (A) Resistance responses of MEA to various concentrations of A549 tumor cells in monoculture for 16 h (at 10mV, 10kHz). (B) This calibration curve represents the correlation between A549 cell density and cell index value. (C) MTS results from of monocultures of A549 cells and MRC-5 fibroblasts after 72 h of exposure to curcumin. The IC50 values for the A549 cells and MRC-5 cells were 50 μM and 30 μM, respectively. The data are shown as the mean (±SD) of 3 repetitions.
Mentions: In the first experimental step, the MEA cell chips were validated via monocultures of A549 tumor cells and MRC-5 fibroblasts in normal proliferative conditions. Before operating the cell measurement system, the arrays were adjusted (by LabVIEW) to normalize the systematic differences, such as wiring, soldering and conductive traces. The normalized cell-free resistance of each electrode was fixed at 2.6 kΩ. Fig 2A shows the resistance responses when single electrodes were challenged by various concentrations of A549 cells. The data obtained over 16 h were fitted by a hill algorithm for a growth/sigmoidal model (OriginPro 9). Below 1.25×105 cells/well, the resistance curves corresponded well to the fitting curves. At higher cell densities, a lag phase appeared from 4 h to 8 h, when the cells were rearranging themselves on binding the site prior to forming a monolayer on the electrodes. To easily estimate viability throughout this study, a dimensionless parameter termed the cell index (CI) was defined as the relative change in measured electrical resistance and can be calculated using the equation belowCI=RiRcell−free−1where Ri is the resistance of the cell-covered electrodes, and Rcell-free is the resistance of blank electrodes (2.6 kΩ). After 16 h, a calibration curve was plotted, which represented the correlation between CI value and cell seeding density (Fig 2B).

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