Esophageal Cancer Cell Culture Dynamics

The esophageal adenocarcinoma cell line, JH-EsoAd157 (link), a kind gift from Dr. James R. Eshleman (The Johns Hopkins University, Baltimore, Maryland, USA), which was previously characterized by Dr.Eshleman, was maintained in complete culture medium containing high-glucose Dulbecco’s Modified Eagle Medium (DMEM with 4 g/L glucose, 4.0 mM L-Glutamine) (Gibco by Life Technologies, Grand Island, NY, USA) supplemented with 10% heat-inactivated FBS (Gibco Life Technologies, Grand Island, NY, USA), and 1% Penicillin-Streptomycin (Gibco by Life Technologies, Grand Island, NY, USA) at 37 °C with 5% CO₂ in microfluidic chips. In the static culture group, the media was changed manually two times per day to minimize the potential effects of hypoxia. In the flow group, adherent cells were continuously supplied with fresh medium using a programmable syringe pump (NewEra Pump Systems Inc., NY) at a flow rate of 2 μl/min. The aldehyde dehydrogenase (ALDH), CD44, CD24 measurements and epithelial to mesenchymal marker stainings were performed on days 1, 3, 5 and 7 to determine the phenotypic plasticity under flow and static conditions.

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Calibasi Kocal G., Güven S., Foygel K., Goldman A., Chen P., Sengupta S., Paulmurugan R., Baskin Y, & Demirci U. (2016). Dynamic Microenvironment Induces Phenotypic Plasticity of Esophageal Cancer Cells Under Flow. Scientific Reports, 6, 38221.

Publication 2016

L-Glutamine heat-inactivated FBS Penicillin-Streptomycin programmable syringe pump

Aldehyde dehydrogenase Cd44 Cell line Cells Chips Eagle Esophageal adenocarcinoma Glucose Glutamine Hypoxia Mesenchymal Penicillin Phenotypic plasticity Stainings Streptomycin Syringe

Corresponding Organization :

Other organizations : Stanford University, Harvard University, Dokuz Eylül University

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Variable analysis

independent variables

Flow condition: The cells were cultured either under continuous flow (2 μl/min) or static conditions.

dependent variables

Aldehyde dehydrogenase (ALDH) expression
CD44 expression
CD24 expression
Epithelial to mesenchymal marker expression

control variables

Cell line: The esophageal adenocarcinoma cell line, JH-EsoAd157, was used.
Culture medium: The cells were maintained in complete culture medium containing high-glucose Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% heat-inactivated FBS and 1% Penicillin-Streptomycin.
Culture conditions: The cells were cultured at 37 °C with 5% CO2.
Measurement timepoints: The measurements were performed on days 1, 3, 5, and 7.

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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