The human hepatoblastoma cell line HepG2 (HB-8065; ATCC, Manassas, VA, USA) was cultured in DMEM-Glutamax™ (5.5 mM glucose) supplemented with 10% fetal bovine serum, streptomycin (100 μg/mL) and penicillin (100 units/mL) at 37 °C in 5% CO2.
Dmem glutamax
Manufactured by American Type Culture Collection
Sourced in United States
DMEM-Glutamax is a powdered cell culture medium formulation provided by American Type Culture Collection (ATCC). It is designed to support the growth and maintenance of a wide range of cell types in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Alamarblue cell viability reagent, by Thermo Fisher Scientific (2 mentions)
Clariostar, by BMG Labtech (2 mentions)
Ku55933, by Bio-Techne (2 mentions)
Dmem glutamax, by Thermo Fisher Scientific (2 mentions)
Parpi, by Selleck Chemicals (2 mentions)
Ht1080 6tg, by American Type Culture Collection (2 mentions)
Dna pkcsi, by Bio-Techne (2 mentions)
Rad51i ri 1, by Selleck Chemicals (2 mentions)
Olaparib, by Selleck Chemicals (2 mentions)
Hacat, by American Type Culture Collection (1 mentions)
Ccl 185, by American Type Culture Collection (1 mentions)
Rpmi 1640 glutamax, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Htb 4, by American Type Culture Collection (1 mentions)
Crl 1573, by American Type Culture Collection (1 mentions)
Penicillin streptomycin pen strep, by Thermo Fisher Scientific (1 mentions)
Crl 2947, by American Type Culture Collection (1 mentions)
Ham s f 12 nutrient mix glutamax, by Thermo Fisher Scientific (1 mentions)
Huh7 cells, by American Type Culture Collection (1 mentions)
Dmem f 12 glutamax, by American Type Culture Collection (1 mentions)
11 protocols using dmem glutamax
1
HepG2 Cell Culture Protocol
2
Evaluating Zinc Chelator Cytotoxicity on HepG2 Cells
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Example 201
Materials and Methods:
Cells
The human hepatoblastoma cell line HepG2 (HB-8065, ATCC, Manassas, Va., USA) was cultured in DMEM-Glutamax™ (5.5 mM glucose) supplemented with 10% foetal bovine serum, streptomycin (100 μg/ml) and penicillin (100 units/ml) at 37° C. in 5% CO2. For viability assays, cells were seeded in white 96-well Nunclon plates at a density of 20,000 cells/well and left overnight to adhere before experiments were conducted.
Cell Viability Assay
Zn chelators dissolved in DMSO were added to white 96-well plates containing 20000 HepG2 cells/well at concentrations ranging from 1 to 1×10−6 mM (max DMSO concentration never exceeded 1%) and incubated for 24 hours at 37° C. After 24 hours, AlamarBlue® cell viability reagent (Thermo Fisher, Carlsbad, USA) was added (10% final concentration) and incubated for 4 hours at 37° C. This dye is a red-ox indicator yielding a fluorescent signal proportional to viable cells in each well that was measured in a fluorescence plate reader at ex550 nm/em603 nm (Clariostar, BMGlabtech, Germany) (O'Brien et al., 2000). Data from 8 replicates were fitted by non-linear regression to estimate IC50 values using GraphPad Prism (GraphPad Software Inc, USA).
3
HepG2 Cell Viability Assay with Zinc Chelators
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Example 201
Materials and methods:
Cells
The human hepatoblastoma cell line HepG2 (HB-8065, ATCC, Manassas, Va., USA) was cultured in DMEM-Glutamax™ (5.5 mM glucose) supplemented with 10% foetal bovine serum, streptomycin (100 μg/ml) and penicillin (100 units/ml) at 37° C. in 5% CO2. For viability assays, cells were seeded in white 96-well Nunclon plates at a density of 20,000 cells/well and left overnight to adhere before experiments were conducted.
Cell Viability Assay
Zn chelators dissolved in DMSO were added to white 96-well plates containing 20000 HepG2 cells/well at concentrations ranging from 1 to 1×10−6 mM (max DMSO concentration never exceeded 1%) and incubated for 24 hours at 37° C. After 24 hours, AlamarBlue® cell viability reagent (Thermo Fisher, Carlsbad, USA) was added (10% final concentration) and incubated for 4 hours at 37° C. This dye is a red-ox indicator yielding a fluorescent signal proportional to viable cells in each well that was measured in a fluorescence plate reader at ex550 nm/em603 nm (Clariostar, BMGlabtech, Germany) (O'Brien et al., 2000). Data from 8 replicates were fitted by non-linear regression to estimate IC50 values using GraphPad Prism (GraphPad Software Inc, USA).
4
Culturing Human Cell Lines
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Human gastric adenocarcinoma (AGS; Sigma-Aldrich), human lung adenocarcinoma (A549; ECACC, Salisbury, UK) and human skin keratinocyte (HaCaT; ATCC, Rockville, MD, USA) cells were maintained with DMEM + GlutaMAX™ supplemented with 10% FBS and 1% penicillin/streptomycin at 37 °C in a humidified atmosphere of 5% CO2.
5
Cell Culture Conditions for Cancer and Kidney Cell Lines
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The human T24 urinary bladder carcinoma cell line (HTB-4; ATCC, Manassas, VA, USA) and the human HEK293 epithelial kidney cell line (CRL-1573; ATCC, Manassas, VA, USA) were cultured in DMEM GlutaMAX. The murine RenCa renal cortical adenocarcinoma cell line from a male BALB/c mouse (CRL-2947, ATCC, Manassas, VA, USA) and the human A549 lung carcinoma cell line (CCL-185, ATCC, Manassas, VA, USA) were cultured in RPMI 1640 GlutaMAX and Ham’s F12 Nutrient Mix GlutaMAX media (31765035, Thermo Scientific™, Waltham, MA, USA), respectively. All media were supplemented with 10% FBS, 100 units/mL of penicillin–streptomycin (pen/strep) (15140122; Thermo Scientific™, Waltham, MA, USA), and 1 mM sodium pyruvate (11360070; Thermo Scientific™, Waltham, MA, USA). All cells were maintained at 37 °C and 5% CO2.
6
Producing and Inactivating HRV16 Stocks
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Human rhinovirus 16 (HRV16) (VR-283, strain 11757, lot 62342987) was purchased from the ATCC and stocks were produced by infecting HeLa Ohio cells in virus medium (DMEM GlutaMax containing 25 mM D-glucose supplemented with 10% FCS and 2 mM l -glutamine) as described previously (Bennett et al, 2012 (link)). Briefly, HeLa Ohio cells were grown to 80% confluence at 37 °C and infected with 5 ml HRV16 or control media for 1 h at room temperature with agitation. The remaining solution was made to 10 ml and the cells with HRV16 left for 48 h at 37 °C to allow for 90% cytopathic effect to develop. Supernatants were then clarified by centrifugation and filtration (Bennett et al, 2012 (link)) and 1 ml stocks were produced and stored at −80 °C. To UV inactivate HRV16 it was treated with UV light (1000 mJ/cm2) for 20 min. Inactivation was confirmed by adding the inactivated virus to HeLa Ohio cells and checking for cytopathic effect.
7
Huh7 Cells Treated with 1-triple TTA
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Human hepatoma Huh7 cells, purchased from ATCC (LGC Standards, Middlesex, UK), were grown in DMEM-Glutamax™ (5.5 mM glucose) medium supplemented with 10% FBS. 1-triple TTA or vehicle (DMSO) were added 48 h before harvesting.
8
Integrin-expressing Cell Lines for Research
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HEK293(β 3 )cells, a human embryonic kidney cell line HEK293 that was stably transfected with the human β 3 integrin gene (kindly provided by J-F. Gourvest, Aventis, France), were cultured in DMEM-GlutaMAX™ supplemented with 10% FBS and 700 μg/mL Geneticin.HEK293(β 3 )-α v RFP cells, a variant cell line of HEK293(β 3 ) transfected with the human α v integrin gene tagged by RFP at its extracellular domain (N-terminus part), were cultured as the HEK293(β 3 ) cells but in the presence of 1 µg/mLPuromycin (See Supplementary information). DU145 cells, a human prostate carcinoma cell line (purchased from ATCC), and M21 cells, a human melanoma cell line, were cultured in DMEM-GlutaMAX™ with 10% FBS. PC3 cells, a human prostate adenocarcinoma cell line (ATCC),were cultured in DMEM-F12-GlutaMAX™ with 10% FBS. Cells were maintained at 37°C, 5% CO 2 in a humidified incubator.
9
Breast cancer cell culture protocol
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Breast cancer cell lines MCF7, MDA-MB-231 and SK-BR-3 were purchased from ATCC and cultured at 37 °C in a humidified atmosphere with 5% CO2 using high-glucose GlutaMAX™ DMEM (MCF-7, MDA-MB-231) and McCoy’s 5A (Modified) Medium (SK-BR-3) supplemented with 10% (v/v) FBS (fetal bovine serum), 1% (v/v) penicillin–streptomycin. Cell lines were characterized by flow cytometry and immunofluorescence for HER2 and HER3 expression (Figure S1 ). For microchip experiments, 10 k cells of each cell line were injected inside Ephesia.
10
Establishing Cell Cultures and Inhibitor Treatments
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HT1080, HT1080 6TG (ATCC) and their derivatives cells were grown in Glutamax-DMEM supplemented with 0.1 mM nonessential amino acids and 10% bovine growth serum. Human IMR-90 primary lung fibroblasts transformed with E6E7 1 (link) were grown in Glutamax-DMEM (Gibco) supplemented with 0.1 mM nonessential amino acids and 15% fetal bovine serum. All cells were grown at 7.5% CO2 and 3% O2. All cell lines were purchased from ATCC or have been commercially authenticated and tested free of mycoplasma. When indicated, cells were treated with the following inhibitors: ATMi (KU-55933, Tocris, 10µM), DNA-PKcsi (NU-7441, Tocris, 1µM), PARPi (Olaparib, Selleck Chemical, 10µM), RAD51i (RI-1, Selleck Chemical, 20µM).
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