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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 84 h of co-culture in the curcumin treatment condition. The raw data were fitted and converted to CI values with respect to the cell-free electrode responses (2.6 kΩ). (B) CI values of the tumor cells at 5 different distances away from the cultured fibroblasts. The inhibitory effect of fibroblasts on tumor cells was more clearly observed in the toxic environment after 24 h (*p <0.04). (C) The migration rate of fibroblasts into the tumor cell area was assessed using a cell tracker staining assay. In the treatment condition, no significant migration was observed. The gray dots indicate the location of the electrodes arrays. The scale bar represents 500 μm.
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pone.0153813.g005: (A) Cellular resistance responses (at 10mV, 10kHz) of A549 tumor cells as a function of time and distance from fibroblasts over 84 h of co-culture in the curcumin treatment condition. The raw data were fitted and converted to CI values with respect to the cell-free electrode responses (2.6 kΩ). (B) CI values of the tumor cells at 5 different distances away from the cultured fibroblasts. The inhibitory effect of fibroblasts on tumor cells was more clearly observed in the toxic environment after 24 h (*p <0.04). (C) The migration rate of fibroblasts into the tumor cell area was assessed using a cell tracker staining assay. In the treatment condition, no significant migration was observed. The gray dots indicate the location of the electrodes arrays. The scale bar represents 500 μm.

Mentions: To provide a full understanding of the neighbor effect between fibroblasts and tumor cells in vitro, we constructed a co-culture model with MRC-5 cells and A549 cells in therapeutic conditions. Curcumin was introduced into the confronted cultures at the previously defined concentration, 30μM; impedance was then monitored for the following 84 h. The average responses were normalized and nonlinearly fitted using a dose-response algorithm (Fig 5A). Interestingly, the distance effect was clearly demonstrated at early times after drug exposure (Fig 5A and 5B). After 24 h, the average CI values obtained were 5.57, 5.77, 6.35, 6.34 and 6.40 for tumor cells that were located at 0.1, 0.25, 0.65, 1.45 and 3.05 mm away from the fibroblast confrontation line. After 48 h and 72 h, significant decreases of all the CI values and the appearance of gaps between the arrays clearly reflected the intensive combined apoptotic effect of both the anti-cancer drug activity and growth inhibition via fibroblast interactions. We also performed a migration assay under the confronted co-culture conditions to investigate the migration ability of fibroblasts into the tumor cells area and thus be able to distinguish the direct cell-to-cell interaction effect from the total inhibitory effect (Fig 5C). After 72 h, the remaining fluorescent intensity showed the efficient apoptotic effect of curcumin on the MRC-5 cells, but no significant migration was observed. These results corresponded well with those of other studies about the effects of curcumin treatment on the migration kinematics of cultured fibroblasts.


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 84 h of co-culture in the curcumin treatment condition. The raw data were fitted and converted to CI values with respect to the cell-free electrode responses (2.6 kΩ). (B) CI values of the tumor cells at 5 different distances away from the cultured fibroblasts. The inhibitory effect of fibroblasts on tumor cells was more clearly observed in the toxic environment after 24 h (*p <0.04). (C) The migration rate of fibroblasts into the tumor cell area was assessed using a cell tracker staining assay. In the treatment condition, no significant migration was observed. The gray dots indicate the location of the electrodes arrays. The scale bar represents 500 μm.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153813.g005: (A) Cellular resistance responses (at 10mV, 10kHz) of A549 tumor cells as a function of time and distance from fibroblasts over 84 h of co-culture in the curcumin treatment condition. The raw data were fitted and converted to CI values with respect to the cell-free electrode responses (2.6 kΩ). (B) CI values of the tumor cells at 5 different distances away from the cultured fibroblasts. The inhibitory effect of fibroblasts on tumor cells was more clearly observed in the toxic environment after 24 h (*p <0.04). (C) The migration rate of fibroblasts into the tumor cell area was assessed using a cell tracker staining assay. In the treatment condition, no significant migration was observed. The gray dots indicate the location of the electrodes arrays. The scale bar represents 500 μm.
Mentions: To provide a full understanding of the neighbor effect between fibroblasts and tumor cells in vitro, we constructed a co-culture model with MRC-5 cells and A549 cells in therapeutic conditions. Curcumin was introduced into the confronted cultures at the previously defined concentration, 30μM; impedance was then monitored for the following 84 h. The average responses were normalized and nonlinearly fitted using a dose-response algorithm (Fig 5A). Interestingly, the distance effect was clearly demonstrated at early times after drug exposure (Fig 5A and 5B). After 24 h, the average CI values obtained were 5.57, 5.77, 6.35, 6.34 and 6.40 for tumor cells that were located at 0.1, 0.25, 0.65, 1.45 and 3.05 mm away from the fibroblast confrontation line. After 48 h and 72 h, significant decreases of all the CI values and the appearance of gaps between the arrays clearly reflected the intensive combined apoptotic effect of both the anti-cancer drug activity and growth inhibition via fibroblast interactions. We also performed a migration assay under the confronted co-culture conditions to investigate the migration ability of fibroblasts into the tumor cells area and thus be able to distinguish the direct cell-to-cell interaction effect from the total inhibitory effect (Fig 5C). After 72 h, the remaining fluorescent intensity showed the efficient apoptotic effect of curcumin on the MRC-5 cells, but no significant migration was observed. These results corresponded well with those of other studies about the effects of curcumin treatment on the migration kinematics of cultured fibroblasts.

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