The 4T1 mammary carcinoma cell line was a gift from Xiao-Fan Wang (Duke University); the cell line was authenticated prior to their use in experiments. 4T1 tumor cells were grown in coordination with ATCC guidelines: ATCC-formulated-RPMI-1640 Medium (ATCC 30–2001) was used and supplemented with 10% fetal bovine serum. Cells were grown at 37°C with 5% CO2 in 100mm cell culture dishes (VWR). Cells were subcultured at 80% confluence at a ratio of 1:6.
100mm cell culture dishes
Manufactured by Avantor
Sourced in United Kingdom
The 100mm cell culture dishes are circular polystyrene dishes designed for the in vitro cultivation of cells. They provide a sterile, controlled environment for the growth and maintenance of various cell lines.
Automatically generated - may contain errors
5 protocols using 100mm cell culture dishes
1
Cell Culture of 4T1 Mammary Carcinoma
2
HEK 293 Cell Lysis and Protein Quantification
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HEK 293 cells to be used as controls in western blots were grown at 37 °C in DMEM plus 10% FBS in 100 mm cell culture dishes (VWR). Cells were collected at 80–90% confluency using RIPA lysis buffer (150 mM NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 50 mM Tris pH8.0) with Roche complete protease inhibitor (Sigma-Aldrich) and HALTTM Phosphatase Inhibitor Cocktail (Thermo Scientific) and normalized to protein concentration measured using Micro BCA Protein Assay Kit (Thermo Scientific).
3
Cell Culture of 4T1 Mammary Carcinoma
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The 4T1 mammary carcinoma cell line was a gift from Xiao-Fan Wang (Duke University); the cell line was authenticated prior to their use in experiments. 4T1 tumor cells were grown in coordination with ATCC guidelines: ATCC-formulated-RPMI-1640 Medium (ATCC 30–2001) was used and supplemented with 10% fetal bovine serum. Cells were grown at 37°C with 5% CO2 in 100mm cell culture dishes (VWR). Cells were subcultured at 80% confluence at a ratio of 1:6.
4
Cell Culture for Gas Exposure
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Cells culture was performed using the same methods described in the in vivo study. Before gas exposure, 0.5 × 105 cells were seeded in 100 mm cell culture dishes (VWR, Leicestershire, UK), followed by incubation at 37 °C and 5% CO2.
5
Sevoflurane Exposure Effects on A549 Cells
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On Day 1, 1 × 105 A549 cells were seeded in 100 mm cell culture dishes (VWR International, Leicestershire, UK) at 37 °C in a 5% CO2 incubator. The A549 cells were exposed to different concentrations of sevoflurane (0%, 2%, 4%) with oxygen for 3 days. Cell culture dishes in a hand-made container with a gas inlet and outlet valves were placed in a 38 °C water bath. Depending on the condition, the different concentrations of sevoflurane and 1 L/min oxygen were delivered to the container. The appropriate concentration of sevoflurane was adjusted by turning the vaporizer dial based on the gas flow analyzer (Datex, Helsinki, Finland). The exposure experiment was conducted three times a week, for 1 h each time. Daily images were obtained by a microscope camera. The confluency of A549 cells reached about 75% on Day 4.
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