HEK293 Cell Culture and Transfection

Human embryonic kidney 293 (HEK293)-derived 293A cells were maintained in Dulbecco’s Modified Eagle’s Medium (Nacalai Tesque, Kyoto, Japan) supplemented with 10% foetal bovine serum (Biowest, Nuaillé, France), and 1% penicillin-streptomycin (Nacalai Tesque), 2 mM L-glutamine (Nacalai Tesque), and 1 × Non-Essential Amino Acid (Life Technologies, Tokyo, Japan) at 37°C in a humidified atmosphere of 5% (v/v) CO₂ in air. Twenty-four hours after the cells had been seeded (0.92 × 10⁵ cells/cm²), each vector plasmid was transfected into 293A cells using polyethyleneimine ‘MAX’ (PEI-MAX) (Polysciences, Warrington, PA, USA) as described previously [23 (link)].

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Toyoda Y., Kawamura Y., Nakayama A., Nakaoka H., Higashino T., Shimizu S., Ooyama H., Morimoto K., Uchida N., Shigesawa R., Takeuchi K., Inoue I., Ichida K., Suzuki H., Shinomiya N., Takada T, & Matsuo H. (2021). Substantial anti-gout effect conferred by common and rare dysfunctional variants of URAT1/SLC22A12. Rheumatology (Oxford, England), 60(11), 5224-5232.

Publication 2021

Dulbecco’s Modified Eagle’s Medium foetal bovine serum penicillin-streptomycin L-glutamine Non-Essential Amino Acid PEI-MAX

Atmosphere Cells Eagle Embryonic Essential amino acid Foetal bovine serum Glutamine Human Kidney Penicillin Plasmid Polyethyleneimine Streptomycin Vector

Corresponding Organization : National Defense Medical College

Other organizations : University of Tokyo Hospital, Sasaki Institute, National Institute of Genetics, Sequenom (United States), Groupe d'Analyse et de Théorie Economique Lyon St Etienne, Nagoya University, Tokyo University of Pharmacy and Life Sciences, Jikei University School of Medicine

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Variable analysis

independent variables

Vector plasmid transfection

dependent variables

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control variables

Dulbecco's Modified Eagle's Medium (Nacalai Tesque, Kyoto, Japan) supplemented with 10% foetal bovine serum (Biowest, Nuaillé, France), and 1% penicillin-streptomycin (Nacalai Tesque), 2 mM L-glutamine (Nacalai Tesque), and 1 × Non-Essential Amino Acid (Life Technologies, Tokyo, Japan)
Incubation at 37°C in a humidified atmosphere of 5% (v/v) CO2 in air
Cell seeding density (0.92 × 10^5 cells/cm^2)

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