Human cell lines (RCC4, HeLa, Hep3B, MCF-7 and U2OS) were cultured in DMEM (D6546-500ML; Sigma Aldrich) each supplemented with 10% fetal bovine serum (F7524-500ML; Sigma Aldrich), 2 mM L-glutamine (G7513- 100ML; Sigma Aldrich), 50 units/ml of penicillin, and 50 μg/ml of streptomycin (P0781-100ML; Sigma Aldrich). The RCC4 cell line was a gift from C.H.C.M. Buys (University of Groningen, [2 (link)]), the HeLa and Hep3B cell lines were from the European Collection of Cell Cultures (ECACC) [2 (link)], the MCF-7 cell line was from the American Type Culture Collection (ATCC) and the U2OS cell line was a gift from S.Galey (ICRF Clare Hall Laboratories, United Kingdom). Cells were treated either with DMSO (control) or compounds (dissolved in DMSO) and added directly into the cell culture medium to the desired final concentration for 4–5 h as previously described [9 (link),22 (link)].
P0781 100ml
Manufactured by Merck Group
Sourced in United States, France
P0781-100ML is a laboratory product offered by Merck Group. It is a 100mL container of the specified product. The core function of this product is to serve as a laboratory reagent or supply, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
G7513 100ml, by Merck Group (2 mentions)
D6546 500ml, by Merck Group (2 mentions)
F7524 500ml, by Merck Group (2 mentions)
Mcf 7 cell line, by American Type Culture Collection (2 mentions)
Hep3b, by Merck Group (1 mentions)
B16f10 cells, by Korean Cell Line Bank (1 mentions)
Trypsin edta 1, by Merck Group (1 mentions)
L glut, by Thermo Fisher Scientific (1 mentions)
D0819 500ml, by Merck Group (1 mentions)
5 protocols using p0781 100ml
1
Culture and Compound Treatment of Human Cell Lines
2
Cell Culture Conditions for MCF-7, Hep3B, and U2OS
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Human cell lines (MCF-7, Hep3B, and U2OS) were cultured in Dulbecco's modified Eagle's medium (DMEM and D6546–500ML; Sigma) each supplemented with 10% fetal bovine serum (F7524–500ML; Sigma), 2 mm l -glutamine (G7513–100ML; Sigma), 50 units/ml penicillin, and 50 μg/ml streptomycin (P0781-100ML; Sigma). The MCF-7 cell line was from the American Type Culture Collection (ATCC); the Hep3B cell line was from the European Collection of Cell Cultures (ECACC) (12 (link)). The U2OS cell line was a gift from S. Galey (ICRF Clare Hall Laboratories, United Kingdom). Cells were treated either with DMSO (control) or compounds (dissolved in DMSO) and added directly into the cell culture medium to the desired final concentration as described previously (27 (link), 35 (link)). For hypoxia (0.5% O2) treatment, cells were seeded at least 24 h prior to being incubated for 16 h in an InvivO2 400 hypoxic workstation (Ruskin Technologies, Bridgend, UK).
3
Murine Melanoma B16F10 and HUVEC Culture
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Murine melanoma B16F10 cells were purchased from the Korean Cell line Bank (80008, Seoul, Korea) and cultured in Dulbecco's modified Eagle's medium (DMEM) (11995065, Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) (EF-0500-A, Atlas Biologicals, For Collins, CO, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin (P0781-100 ML, Sigma-Aldrich, St. Louis, MO, USA). Human umbilical vein endothelial cells (HUVECs) and their growth medium MV2 were purchased from Promo Cell (C-22010, Heidelberg, Germany). The cells were used at passages 4-5 in all experiments. B16F10 cells and HUVECs were incubated at 37°C with 5% CO2. Methyl gallate (MG, PHL82592, Sigma-Aldrich) was dissolved in dimethyl sulfoxide (DMSO) to obtain a stock solution at 500 mM (for in vitro use) or 4 mg/ml (for in vivo use), which was then diluted with autoclaved PBS buffer.
4
Preparing Cells for Single-Cell Experiments
5
Murine Macrophage RAW 264.7 Cell Culture
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The murine macrophage cell line RAW 264.7 (TIB-71, ATCC) (originating from BALB/c mice cells transformed with Abelson leukaemia virus [43 (link)]) was used as a model for the induction of a pro-immune or anti-inflammatory response. For facility of passaging, cells were cultured normally as a semi-adherent culture in 10 cm2 suspension culture dishes at 37 °C, 5% CO2 and 80% humidity in 12 mL DMEM (#D0819-500ML, Sigma-Aldrich, Saint-Quentin-Fallavier, France) medium supplemented with 5% FBS 100 U/mL penicillin and 0.1 mg/mL streptomycin (#P0781-100ML, Sigma). Cells were passaged by gentle flushing of the culture medium to remove the cells, with cells then pelleted (200 g, 3 min, RT), resuspended in medium and counted (Section 2.2.1 ). Cells were re-seeded at 5 × 106 cells/mL or 10 × 106 cells/mL for 2 or 3 days of growth, respectively.
For experiments, RAW 264.7 cells were seeded the day before treatment at a density of 3 × 104 cells per well (100µL medium volume) in regular adherent 96-well culture plates (#655180, Greiner Bio-One, Les Ulis, France) or 3 × 105 cells per well (300µL medium volume) in 12-well plates (#665180, Greiner Bio-One, Les Ulis, France).
For experiments, RAW 264.7 cells were seeded the day before treatment at a density of 3 × 104 cells per well (100µL medium volume) in regular adherent 96-well culture plates (#655180, Greiner Bio-One, Les Ulis, France) or 3 × 105 cells per well (300µL medium volume) in 12-well plates (#665180, Greiner Bio-One, Les Ulis, France).
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