Gallium(III) meso-tetraphenylporphyrin chloride (GaTP) was purchased from Frontier Scientific (Logan, UT). THP-1 cells were purchased from ATCC. Difco Middlebrook 7H9 broth and BBL Middlebrook oleic acid-albumin-dextrose-catalase (OADC) enrichment medium were purchased from BD (Sparks, MD, USA). 7H10 agar plates were purchased from Remel Inc. (Lenexa, KS). HyClone RPMI 1640, Dulbecco modified Eagle medium (DMEM), l -glutamine, HEPES, sodium pyruvate, and fetal bovine serum were purchased from GE Life Sciences (Logan, UT). Macrophage colony-stimulating factor (MCSF) was purchased from BioLegend (San Diego, CA). Fe-free 7H9 medium was prepared as described previously (11 (link)). Human monocytes that had been purified by countercurrent centrifugal elutriation from normal human donors were purchased from the UNMC Elutriation core facility using an institutional IRB-approved protocol. HIV-1ADA was obtained from UNMC, Omaha, and its potency was tested by reverse transcriptase (RT) assay before infection. H37Rv was obtained from ATCC and grown in a biosafety level 3 (BSL3) lab, and its potency was tested by counting CFU before infection.
Sodium pyruvate
Manufactured by GE Healthcare
Sourced in United States, Germany, United Kingdom, Austria
Sodium pyruvate is a chemical compound that serves as an energy substrate in cell culture media. It is commonly used in laboratory settings to support the growth and metabolism of cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Non essential amino acids (neaa), by GE Healthcare (13 mentions)
Penicillin, by Thermo Fisher Scientific (11 mentions)
Streptomycin, by Thermo Fisher Scientific (11 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
β mercaptoethanol, by Merck Group (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Hepes, by GE Healthcare (5 mentions)
Penicillin, by Gemini Bio (5 mentions)
Streptomycin, by Gemini Bio (5 mentions)
Dulbecco modified eagle medium (dmem), by Corning (5 mentions)
Hepes, by Corning (5 mentions)
L glutamine, by GE Healthcare (5 mentions)
L glutamine, by Thermo Fisher Scientific (5 mentions)
Glutamax, by Thermo Fisher Scientific (5 mentions)
Trypsin edta, by Thermo Fisher Scientific (4 mentions)
Gentamycin, by Avantor (4 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (4 mentions)
Non essential amino acid (neaa), by Lonza (3 mentions)
Fetal bovine serum (fbs), by Gemini Bio (3 mentions)
Penicillin streptomycin, by GE Healthcare (3 mentions)
49 protocols using sodium pyruvate
1
Gallium(III) Porphyrin Antitubercular Evaluation
2
MNK-3 Cell Culture Maintenance Protocol
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MNK-3 cell line [45 (link)] and clone B3 were maintained in DMEM (Corning; Tewksbury, MA) with 10% heat-inactivated FBS (Gemini Bio-Products; West Sacramento, CA), 2 mM GlutaGro (Corning), 1 mM sodium pyruvate (GE Healthcare HyClone; Logan, UT), 55 μM β-mercaptoethanol (Sigma), 10 mM HEPES (Corning), 50 μg/ml gentamycin (Amresco; Solon, OH), 100 U/ml penicillin (Gemini Bio-Products), 100 U/ml streptomycin (Gemini Bio-Products) and 10 ng/ml recombinant mouse IL-7 and IL-15 (eBioscience; San Diego, CA or Peprotech; Rocky Hill, NJ).
3
Culturing and Implanting Tumor Cell Lines
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B16-F10, TC-1, and B16.OVA were cultured in RPMI-1640 media (GE Life
Sciences), and MC38 was cultured in DMEM media (Gibco), each supplemented with
10% v/v heat-inactivated FCS (Atlanta Biologicals), 100 U/mL penicillin, 100
μg/mL streptomycin (Gibco), 1× non-essential amino acids (GE Life
Sciences), and 1 mM sodium pyruvate (GE Life Sciences). B16.OVA media was
supplemented with 0.5 mg/mL G418. For each tumor cell implantation, a frozen
aliquot was thawed, passaged once, and harvested using trypsin EDTA (Gibco),
quenched with HI-FCS, washed in PBS, and implanted subcutaneously in sterile
PBS. Tumors were measured using digital calipers twice per week, and tumor
volume was estimated using the formula [tumor volume = short × short
× long / 2]. Animals were euthanized when tumors reached size criteria
(1000 mm3 or 2000 mm3).
Sciences), and MC38 was cultured in DMEM media (Gibco), each supplemented with
10% v/v heat-inactivated FCS (Atlanta Biologicals), 100 U/mL penicillin, 100
μg/mL streptomycin (Gibco), 1× non-essential amino acids (GE Life
Sciences), and 1 mM sodium pyruvate (GE Life Sciences). B16.OVA media was
supplemented with 0.5 mg/mL G418. For each tumor cell implantation, a frozen
aliquot was thawed, passaged once, and harvested using trypsin EDTA (Gibco),
quenched with HI-FCS, washed in PBS, and implanted subcutaneously in sterile
PBS. Tumors were measured using digital calipers twice per week, and tumor
volume was estimated using the formula [tumor volume = short × short
× long / 2]. Animals were euthanized when tumors reached size criteria
(1000 mm3 or 2000 mm3).
4
Virus Propagation in MRC-5 Fibroblasts
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For virus propagation, MRC-5 pulmonary fibroblasts (ECACC, ref. 05090501) were grown in antibiotic-free Minimum Essential Media (MEM) with nonessential amino acids (Thermo Fisher Scientific), complemented with 10% foetal bovine serum (FBS, Eurobio), 2 mM glutamine (Sigma Aldrich), 1% nonessential amino acids (Gibco), and 1% sodium pyruvate (GE Healthcare). For the antiviral assays, the same medium was used, containing only 2% FBS.
5
HepG2 Cell Culture Methodology
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The HepG2 cell line was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA), and the cells were resuspended in DMEM supplemented with 10% FBS (Gibco BRL Co., Gaithersburg, MD, USA), 1% antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin) (Gibco BRL Co., Gaithersburg, MD, USA), 1% sodium pyruvate (Hyclone, GE Healthcare, Pittsburge, PA, USA), and 2 mM glutamine (Gibco BRL Co., Gaithersburg, MD, USA) at 37 °C in a humidified atmosphere with 5% CO2.
6
Culturing Immortalized Human Brain Microvascular Endothelial Cells
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Immortalized human brain microvascular endothelial cells (HBMEC) were kindly provided by K. S. Kim (Stins et al., 1997 (link)) and were cultured as described previously (Unkmeir et al., 2002 (link)). Briefly, cells were cultured in RPMI-1640 medium supplemented with 1% sodium pyruvate (1 mM), 1% L-glutamine (2 mM), 1% non-essential amino acids (all purchased from GE Healthcare, Little Chalfont, United Kingdom), 5 U/ml heparin (Biochrom, Berlin, Germany) and 30 μg/ml endothelial cell growth supplement (ECGS, CellSystems, Troisdorf, Germany). Cells were incubated at 37°C and 5% CO2 in a humidified atmosphere.
7
Cytokine Production in Thymocytes and Splenocytes
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Five million thymocytes or splenocytes were resuspended in 3 ml of complete RPMI1640 medium [Sigma, supplemented with 10% FCS (Sigma), 100 U/ml penicillin–streptomycin (GE Healthcare), 2 mM l -glutamine (Sigma), 0.1 mM non-essential amino acid (Lonza), 1 mM sodium pyruvate (GE Healthcare), 55 μM β-mercaptoethanol (Sigma)] containing 50 ng/ml of phorbol 12-myristate 13-acetate (PMA; Sigma) and 500 ng/ml of ionomycin (Sigma) in the presence of Golgistop (BD Biosciences). Cells were harvested after 4.5 h incubation at 37 °C, and were stained with CD1d-tet as well as appropriate antibodies for surface markers. Cells were fixed with BD fixation buffer (BD Biosciences), and were stained for intracellular cytokines in 1 × BD perm buffer (BD Biosciences). Cytokine production was measured by LSRII or Fortessa (BD Biosciences).
8
TGFβ SMAD Reporter Assay in HEK293 Cells
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TGFβ SMAD reporter assay was performed as per the manufacturer's instructions (BPS Biosciences, catalog no. 60653). Briefly, HEK293 cells were plated at a density of approximately 35,000 cells per well on to 96-well white clear-bottom plates in growth media containing MEM/EBSS (HyClone Laboratories Inc., GE Healthcare Lifesciences) with 1% non-essential amino acids (HyClone Laboratories Inc., GE Healthcare Lifesciences), 1 mmol/L sodium pyruvate (HyClone Laboratories Inc., GE Healthcare Lifesciences), 1% pen-strep (GIBCO, Thermo Fisher Scientific), and 10% FBS (MP Bio Science Ltd) and allowed to attach overnight. Spent media was discarded and cells were then incubated with recombinant TGFβ (BPS Biosciences) at 20 ng/mL and with different dilutions of BCA101, human IgG isotype antibody (R&D Systems Inc.) or TGFβRII-Fc at equimolar concentration prepared in assay media (growth media containing 0.5% FBS). After 18 hours, cells were incubated with 100 μL of BioGlo luciferase reagent for 15 minutes at room temperature. Luminescence readout was taken as per manufacturer's instructions using a Spectramax M5e plate reader (Molecular Devices) with SoftMax Pro GxP software version 6.5. EC50 was analyzed using SoftMax Pro software and data plotted using GraphPad Prism software version 9.
9
Caco-2 Colorectal Cancer Cell Culture
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Colorectal cancer cell line, Caco-2, was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Caco-2 cells were cultured in normal medium: DMEM (Dulbecco’s Modified Eagle Medium, GIBCO, Thermo Fisher Scientific Inc., Waltham, MA, USA), high glucose medium, containing 20% FBS (Fetal bovine serum, Thermo Fisher Scientific Inc.), 0.02 mM L-Glutamine (Thermo Fisher Scientific Inc.), 0.1× non-essential amino acid (Thermo Fisher Scientific Inc.), 0.01 mM sodium pyruvate (Hyclone, GE Healthcare, Pittsburge, PA, USA), and 1.5 g/L sodium bicarbonate (Sigma, St. Louis, MO, USA).
10
HepG2 Cell Line Maintenance Protocol
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The human hepatoblastoma cell line, HepG2 (22 (link)) was maintained in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA), supplemented with 10% heat-inactivated bovine serum (Gibco; Thermo Fisher Scientific, Inc.), 1X penicillin-streptomycin (Gibco; Thermo Fisher Scientific, Inc.) and 1X sodium pyruvate (GE Healthcare Life Sciences, Logan, UT, USA) at 37°C in a humidified chamber containing 5% CO2. Silymarin, 2,7-dichlorofluorescein (DCFH), ascorbic acid (all Sigma-Aldrich; Merck KGaA) and gallic acid (Fluka; Honeywell International Inc., Morris Plains, NJ, USA) were also purchased.
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