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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) Cellular resistance responses (at 10mV, 10kHz) of A549 tumor cells as a function of time and distance from fibroblasts over 54 h of co-culture in normal growth conditions. The raw data were linearly-fitted and converted to CI value with respect to the cell-free electrode response (2.6 kΩ). (B) CI values of tumor cells at 5 different distances away from the cultured fibroblasts. After 48 h in co-culture, the CI values indicated that the tumor cells that were confronted with fibroblasts were clearly inhibited in comparison with distant cells. (*p < 0.02). (C) Bright-field images of the confrontation between tumor cells and fibroblasts at 0 h and 48 h. The scale bar represents 200 μm.
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pone.0153813.g004: (A) Cellular resistance responses (at 10mV, 10kHz) of A549 tumor cells as a function of time and distance from fibroblasts over 54 h of co-culture in normal growth conditions. The raw data were linearly-fitted and converted to CI value with respect to the cell-free electrode response (2.6 kΩ). (B) CI values of tumor cells at 5 different distances away from the cultured fibroblasts. After 48 h in co-culture, the CI values indicated that the tumor cells that were confronted with fibroblasts were clearly inhibited in comparison with distant cells. (*p < 0.02). (C) Bright-field images of the confrontation between tumor cells and fibroblasts at 0 h and 48 h. The scale bar represents 200 μm.

Mentions: To discriminate the particular contribution of each cell line to the total inhibitory effect, we examined cellular conditions via impedance responses in a co-culture model. Raw resistance data of A549 cells at various distances away from fibroblasts (0.1, 0.25, 0.65, 1.45, and 3.05 mm) were linearly fitted and normalized, as shown in Fig 4A. The corresponding resistance data after 24 h and 48 h were converted into CI values using the equation shown in section 1 (Fig 4B). During the first 24 h after cell contact, the distance effect was indistinct, and the CI values did not show significant differences between the 1st electrode array (0.1 mm) and the 5th electrode array (3.05 mm).


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) Cellular resistance responses (at 10mV, 10kHz) of A549 tumor cells as a function of time and distance from fibroblasts over 54 h of co-culture in normal growth conditions. The raw data were linearly-fitted and converted to CI value with respect to the cell-free electrode response (2.6 kΩ). (B) CI values of tumor cells at 5 different distances away from the cultured fibroblasts. After 48 h in co-culture, the CI values indicated that the tumor cells that were confronted with fibroblasts were clearly inhibited in comparison with distant cells. (*p < 0.02). (C) Bright-field images of the confrontation between tumor cells and fibroblasts at 0 h and 48 h. The scale bar represents 200 μm.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153813.g004: (A) Cellular resistance responses (at 10mV, 10kHz) of A549 tumor cells as a function of time and distance from fibroblasts over 54 h of co-culture in normal growth conditions. The raw data were linearly-fitted and converted to CI value with respect to the cell-free electrode response (2.6 kΩ). (B) CI values of tumor cells at 5 different distances away from the cultured fibroblasts. After 48 h in co-culture, the CI values indicated that the tumor cells that were confronted with fibroblasts were clearly inhibited in comparison with distant cells. (*p < 0.02). (C) Bright-field images of the confrontation between tumor cells and fibroblasts at 0 h and 48 h. The scale bar represents 200 μm.
Mentions: To discriminate the particular contribution of each cell line to the total inhibitory effect, we examined cellular conditions via impedance responses in a co-culture model. Raw resistance data of A549 cells at various distances away from fibroblasts (0.1, 0.25, 0.65, 1.45, and 3.05 mm) were linearly fitted and normalized, as shown in Fig 4A. The corresponding resistance data after 24 h and 48 h were converted into CI values using the equation shown in section 1 (Fig 4B). During the first 24 h after cell contact, the distance effect was indistinct, and the CI values did not show significant differences between the 1st electrode array (0.1 mm) and the 5th electrode array (3.05 mm).

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