Culturing Renal Cell Lines

Human renal cancer 786-O and ACHN cell lines were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan) and cultured in RPMI-1640 medium (Cytiva, SH30011.02) and MEM medium (Cytiva, SH30193.04). The human proximal tubule epithelial HK2 cell line was incubated with DMEM/F12 medium (Cytiva SH30004.03). These cell culture media contained 10% FBS (Cytiva SH30071.02), NEAA solution (100X; Cytiva SH30238.01), P/S reagent (100X, Cytiva SV30010) and cells incubated at 37 °C (involved in 5% CO₂) in a humidified atmosphere. RCC and HK-2 cells were subcultured when cells reached 70% confluence.

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Tseng T.Y., Lee C.H., Lee H.L., Su C.Y., Kao C.Y., Tsai J.P, & Hsieh Y.H. (2023). Licochalcone A Suppresses Renal Cancer Cell Proliferation and Metastasis by Engagement of Sp1-Mediated LC3 Expression. Pharmaceutics, 15(2), 684.

Publication 2023

MEM medium DMEM/F12 medium

Atmosphere Cell culture Cell lines Cells Culture media Epithelial cell Human Proximal tubule Renal cancer

Corresponding Organization : Tzu Chi University

Other organizations : China Medical University, National Yang Ming Chiao Tung University

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Variable analysis

independent variables

Cell lines used: 786-O, ACHN, and HK2

dependent variables

Not explicitly mentioned

control variables

Cell culture media: RPMI-1640, MEM, and DMEM/F12
Supplements: 10% FBS, NEAA solution, and P/S reagent
Incubation conditions: 37°C, 5% CO2, and humidified atmosphere
Subculturing when cells reached 70% confluence

controls

No positive or negative controls were explicitly mentioned in the provided information.

Annotations

Based on most similar protocols

Maintain the cell culture passage number below 30 for Caki-1 and not more than 40 for ACHN cells to ensure consistent experimental performance (Protocol 1).

Validate the cell lines by confirming their identity and checking for mycoplasma contamination to ensure the accuracy of the results (Protocols 1-5).

Use FBS (10%) and P/S (1%) supplements in the cell culture media to support cell growth and maintain cell viability (Protocols 1-5).

Incubate the cells at 37°C in a humidified atmosphere with 5% CO2 to mimic the physiological conditions and optimize cell growth (Protocols 1-5).

Subculture the cells when they reach 70% confluence to maintain the cells in the exponential growth phase and ensure reproducibility (Input Protocol).

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