Sodium pyruvate

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Sodium pyruvate is a chemical compound commonly used in cell culture media. It serves as an energy source for cells and is involved in various metabolic processes. Sodium pyruvate is a key intermediate in the citric acid cycle, which is the central pathway for cellular respiration and energy production.

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Pyruvate (379 citations) Sodium pyruvate solution (34 citations) MEM sodium pyruvate (29 citations) Sodium Pyruvate 100 mM Solution (19 citations) Insulin–transferrin–selenium–sodium pyruvate (11 citations) HEPES and sodium pyruvate (4 citations) DMEM without sodium pyruvate (2 citations) Intraperitoneal sodium pyruvate (2 citations) Glucose and Sodium Pyruvate (2 citations) Sodium pyruvate 100× (2 citations)

4 375 protocols using sodium pyruvate

Isolation and Culture of Mouse Embryonic Fibroblasts and Cell Lines

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Mouse embryonic fibroblasts (MEFs) were isolated from 13.5 d.p.c C57BL/6 mouse embryos. MEFs were cultured in DMEM/high glucose (HyClone), 10% FBS (Natocor) supplemented with 1% non‐essential amino acids (NEAA; Gibco). PSCs were maintained on feeder layers (MEFs) in DMEM/high glucose medium supplemented with 15% FBS (Gibco), 1% NEAA (Gibco), 1% GlutaMAX (Gibco), 1% sodium pyruvate (Gibco), 0.1 mM β‐mercaptoethanol (Sigma), 1 μM PD0325901 (Selleck), 3 μM CHIR‐99021 (Selleck), and 1000 U/ml LIF (Peprotech). E.G7‐OVA cell line (ATCC) was cultured in RPMI‐1640 (Gibco) supplemented with 10% FBS (Natocor), 1% GlutaMAX (Gibco), 1% sodium pyruvate (Gibco), and 0.1 mM β‐mercaptoethanol (Sigma). B16F10 melanoma cell line was purchased from the Cell Bank of Chinese Academy of Sciences and stably transduced with chicken OVA cDNA to generate B16F10‐OVA cell line.²³ B16F10‐OVA cells were cultured in complete medium [RPMI‐1640 (Gibco) supplemented with 10% FBS (Natocor), 1% sodium pyruvate (Gibco), and 1% GlutaMAX (Gibco)]. OP9‐DL1 cells were cultured in α‐MEM supplemented with 15% FBS and 1% GlutaMAX (Gibco). The AFT024 cell line (ATCC) was maintained in DMEM/high glucose and 10% FBS (Natocor) supplemented with 0.1 mM β‐mercaptoethanol (Sigma) and 1% sodium pyruvate (Gibco).

Wu B., Zhang Q., Hong P., Liu L., Peng H., Xia C., Wang T., Wang Y., Weng Q., Liu X., Geng Y., Wang J, & Wu H. (2023). Antigen‐specific TCR‐T cells from Rag2 gene‐deleted pluripotent stem cells impede solid tumour growth in a mouse model. Cell Proliferation, 56(4), e13389.

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Mammalian Cell Culture Conditions

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HeLa, COS-7 and HEK cells were cultured at 37°C and 5% CO₂ in DMEM medium (Life Technology) containing 10% fetal calf serum, 1 mM sodium pyruvate, 100 U/ml penicillin, 100 μg/ml streptomycin and 2 mM glutamax (Life Technology). N2A and PC12 cells were cultured in RPMI containing 10% fetal calf serum, 1 mM sodium pyruvate, 100 U/ml penicillin, 100 μg/ml streptomycin and 2 mM glutamax (Life Technology). INS1 cells (passage 30 to 50) were plated in RPMI-1640 with 11.1 mM D-Glucose and supplemented with 10% fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 10 mM HEPES-Na pH 7.4, 2 mM L-glutamine, 1 mM sodium pyruvate (Life Technologies), and 50 μM β-mercaptoethanol (Sigma Aldrich). 18 hours before experiments the standard medium with 11.1 mM glucose was replaced with fresh medium containing 5 mM glucose.
In all our experiments, plasmids were transfected by using Lipofectamine 2000 (Life Technology) and cells were cultured for 24 hours before analysis.

Lees J.A., Messa M., Sun E.W., Wheeler H., Torta F., Wenk M.R., De Camilli P, & Reinisch K.M. (2017). Lipid transport by TMEM24 at ER–plasma membrane contacts regulates pulsatile insulin secretion. Science (New York, N.Y.), 355(6326), eaah6171.

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Cell culture maintenance and viability

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HCT116 (ATCC), HT-29 (ECACC) were maintained in RPMI 1640 (Gibco) supplemented with 10% FBS (Gibco), 1% Penicillin -Streptomycin, 1% non-essential amino acids (Gibco), 1 mM Sodium pyruvate (Gibco) and 10 mM HEPES (Sigma) in a 5% CO₂ incubator. Thp1 XBlue, HEK Null1, TLR2, TLR4, NOD1, and NOD2 bearing NFkB-AP1 reporter (Invivogen) were maintained in the same conditions. Caco-2 (ATCC) were cultured in DMEM supplemented with 10% FBS (Difco), 1% Penicillin-Streptomycin (Gibco), 1% MEM non-essential amino acids (Gibco), 1 mM Sodium pyruvate (Gibco), and 10 mM HEPES (Sigma) in a 10% CO₂. T84 (ATCC) were grown in DMEM-F12 medium supplemented with 10% FBS (Gibco), 1% Penicillin-Streptomycin (Gibco), 1% MEM non-essential amino acids (Gibco), 1 mM Sodium pyruvate (Gibco), and 10 mM HEPES (Sigma) in a 5% CO₂ incubator. Mycoplasma and bacterial contaminations were tested regularly by PCR or using HEK TLR2 cell-line. Cell viability was monitored by MTS measurement using the CellTiter 96 Aqueous One solution (Promega) according to the manufacturer’s recommendations.

Martin-Gallausiaux C., Malabirade A., Habier J, & Wilmes P. (2020). Fusobacterium nucleatum Extracellular Vesicles Modulate Gut Epithelial Cell Innate Immunity via FomA and TLR2. Frontiers in Immunology, 11, 583644.

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Cell Culture Conditions for MLPS and MSC

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Three MLPS cell lines, two MSC lines, and a HEK293T LentiX cell line were used in this study. The MLPS-1765-92 and MLPS-402-91 cell lines were generous gifts from Pierre Åman (University of Gothenburg, Sweden) and the MLPS-DL221 cell line was a generous gift from Alexander Lazar (MD Anderson). The hTERT-MSC line (ASC52telo, hTERT immortalized adipose derived Mesenchymal stem cells, ATCC^® SCRC-4000) was purchased from ATCC.
MLPS-1765-92 and MLPS-402-91 cells were cultured in RPMI medium (Gibco) supplemented with 10% fetal bovine serum, 1% Glutamax (Gibco), 1% Sodium Pyruvate (Gibco), and 1% Penicillin-Streptomycin (Gibco). MLPS-DL221 cells were cultured in DMEM medium (Gibco) supplemented with 10% fetal bovine serum, 1% Glutamax (Gibco), 1% Sodium Pyruvate (Gibco), and 1% Penicillin-Streptomycin (Gibco). hTERT-MSC and AD-MSC lines were cultured in Mesenchymal Stem Cell Basal Medium (ATCC No. PCS-500-030) supplemented with the Mesenchymal Stem Cell Growth Kit for Adipose and Umbilical-derived MSCs - Low Serum (ATCC No. PCS-500-040) and 0.1% Penicillin-Streptomycin (Gibco). HEK293T LentiX cells were cultured in DMEM medium (Gibco) supplemented with 10% fetal bovine serum, 1% Glutamax (Gibco), 1% Sodium Pyruvate (Gibco), 1% HEPES (Gibco), 1% MEM NEAA (Gibco), and 1% Penicillin-Streptomycin (Gibco). All cells were maintained in a humidified incubator at 37°C with 5% CO2.

Zullow H.J., Sankar A., Ingram D.R., Guerra D.D., D’Avino A.R., Collings C.K., Segura R.N., Yang W.L., Liang Y., Qi J., Lazar A, & Kadoch C. (2022). The FUS::DDIT3 fusion oncoprotein inhibits BAF complex targeting and activity in myxoid liposarcoma. Molecular cell, 82(9), 1737-1750.e8.

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Isolation and Culture of Single Myofibers

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Single myofibers were isolated from FDB muscles of 6–8 week old Myf5-Cre/R26R-eYFP mice (adapted from EDL myofiber isolations used by Shefer and Yablonka-Reuveni, 2005 (link)) following dissection and incubation in DMEM with 2% L-glutamine, 4.5% glucose, and 110 mg/mL sodium pyruvate (Gibco) containing 0.2% collagenase I (Sigma) for 2 hours at 37°C with agitation. Myofibers were isolated using gentle trituration in DMEM+ with 2% L-glutamine, 4.5% glucose, and 110 mg/ml sodium pyruvate (Gibco) containing 20% FBS (Wisent) with a glass pipet. and cultured at 37°C in suspension in 96-well dishes containing DMEM+ with 2% L-glutamine, 4.5% glucose, and 110 mg/ml sodium pyruvate (Gibco) containing 20% FBS (Wisent) and 1% chick embryo extract (CEE, Accurate Chemicals) and supplemented with either 1:1000-dilution of DMSO (Sigma), DMSO +Wnt7a (50ng/mL; R&D Systems), or 1μM of a small molecule compound. Fibers were collected after 42h of culture and genomic DNA was isolated and purified using the Dneasy 96-well Blood and Tissue kit (Qiagen). qPCR enumeration of eYFP^Pos and eYFP^Neg cells was performed as described above. Primary screening was performed in biological duplicates, each with technical duplicate qPCR reactions. Results were normalized to DMSO-only controls and averaged between replicates.

Wang Y.X., Feige P., Brun C.E., Hekmatnejad B., Dumont N.A., Renaud J.M., Faulkes S., Guindon D, & Rudnicki M.A. (2019). EGFR-Aurka Signaling Rescues Polarity and Regeneration Defects in Dystrophin-Deficient Muscle Stem Cells by Increasing Asymmetric Divisions. Cell stem cell, 24(3), 419-432.e6.

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Maintenance of Diverse Cell Lines

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HEK293T cells were obtained from the American Type Culture Collection (ATCC) and maintained in Dulbecco’s modified Eagle Medium (DMEM, GIBCO) with 10% fetal bovine serum (FBS, Sigma Aldrich), 1x Penicillin/Streptomycin (GIBCO), 1 mM Sodium Pyruvate (GIBCO), 2 mM L-Glutamine (Thermo Fisher Scientific) at 37°C/5% CO₂. The HeLa-derived TZM-bl reporter cell line was sourced from the NIH AIDS Reagent Program and maintained in DMEM containing 10% fetal bovine serum, 1 mM Sodium Pyruvate, 2 mM L-Glutamine (Thermo Fisher Scientific), 50 μg/ml Gentamicin (Sigma-Aldrich), and 25 mM HEPES (Biochrom) at 37°C/5% CO₂. HEK293EBNA1-6E (293-6E) cells were obtained from the National Research Council Canada (NRC) and maintained in Freestyle 293 Expression Medium (Thermo Fisher Scientific) containing 0.2% Penicillin/Streptomycin at 37°C /5% CO₂ with shaking at 90-120 rpm. The sex of these cell lines is unknown. CHO Flp-In™ cells (Invitrogen) were a kind gift from the lab of John Moore (Cornell University) and maintained in Ham’s F-12 Medium supplemented with 10% heat-inactivated FBS (Sigma-Aldrich), 200 U/ml Penicillin/Streptomycin, 2 mM L-Glutamine, 20 mM HEPES, 0.1 mM non-essential amino acids, 1 mM Sodium Pyruvate (GIBCO), and further supplemented with 100 μg/ml Zeocin (Invitrogen).

Schoofs T., Barnes C.O., Suh-Toma N., Golijanin J., Schommers P., Gruell H., West AP J.r., Bach F., Lee Y.E., Nogueira L., Georgiev I.S., Bailer R.T., Czartoski J., Mascola J.R., Seaman M.S., McElrath M.J., Doria-Rose N.A., Klein F., Nussenzweig M.C, & Bjorkman P.J. (2019). Broad and Potent Neutralizing Antibodies Recognize the Silent Face of the HIV Envelope. Immunity, 50(6), 1513-1529.e9.

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Cell Culture Protocols for Various Cell Types

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HEK293 and HeLa cells were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMED; Gibco) supplemented with 10% FBS (Hyclone) and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin; Gibco). For MEFs, the cell culture medium consists of 10% FBS (Hyclone), 1 mM sodium pyruvate (Gibco), 0.055 mM β-mercaptoethanol (Gibco), 2 mM L-GlutaMax (Gibco), 0.1 mM NEAA (Gibco), and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin; Gibco). For iPSC generation cell culture media, 1000 U/ml LIF (Millipore) were added based on MEF media. iPSC stable clones were maintained on mitomycin C-treated MEF feeder cells in knockout DMEM medium (Gibco) supplemented with 15% KSR, 1 mM sodium pyruvate (Gibco), 0.055 mM β-mercaptoethanol (Gibco), 2 mM L-GlutaMax (Gibco), 0.1 mM NEAA (Gibco), 100 U/ml penicillin (Gibco), 100 μg/ml streptomycin (Gibco) and 1000 U/ml LIF (Millipore). CGR8 ES cells were cultured on gelatin-coated dishes, in GMEM medium supplemented with 10% FBS (Millipore), 1 mM sodium pyruvate (Gibco), 0.055 mM β-mercaptoethanol (Gibco), 2 mM L-GlutaMax (Gibco), 0.1 mM NEAA (Gibco), 100 U/ml penicillin (Gibco), 100 μg/ml streptomycin (Gibco) and 1000 U/ml LIF (Millipore). P19 EC cells were maintained on gelatin-coated dishes, in α-MEM medium supplemented with 10% FBS (Hyclone) and 100 U/ml penicillin (Gibco), 100 μg/ml streptomycin (Gibco).

Jin W., Wang L., Zhu F., Tan W., Lin W., Chen D., Sun Q, & Xia Z. (2016). Critical POU domain residues confer Oct4 uniqueness in somatic cell reprogramming. Scientific Reports, 6, 20818.

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Pkd1 Knockout MEF Proliferation

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Immortalized Pkd1^+/+ and Pkd1^−/− MEFs were plated at a density of 150,000 cells/well in DMEM (Gibco) supplemented with 0.5% FBS (Gibco) and 1% Pen/Strep (Gibco). After 16 h, medium was changed to control medium (DMEM, Gibco), supplemented with 10% FBS (Gibco), 1% Pen/Strep (Gibco), sodium pyruvate (1 mM; Gibco), sodium bicarbonate (44 mM; Sigma Aldrich), 25 mM glucose (Sigma Aldrich), and 4 mM glutamine (Gibco); glucose starvation medium (DMEM supplemented with 1 mM sodium pyruvate, 44 mM sodium bicarbonate, 10% FBS, 1% Pen/Strep and 4 mM glutamine); glutamine starvation medium (DMEM supplemented with 10% FBS, 1% Pen/Strep, 1 mM sodium pyruvate, 44 mM sodium bicarbonate, and 25 mM glucose) or glucose and glutamine starvation medium (DMEM supplemented with 10% FBS and 1% Pen/Strep, 1 mM sodium pyruvate, 44 mM sodium bicarbonate). After 24 and 48 h, cells were trypsinized and counted with an automated cell counter (Countess cell counter, Invitrogen). Pictures from each sample at both time points were taken with a white field microscope, using a ×10 objective.

Podrini C., Rowe I., Pagliarini R., Costa A.S., Chiaravalli M., Di Meo I., Kim H., Distefano G., Tiranti V., Qian F., di Bernardo D., Frezza C, & Boletta A. (2018). Dissection of metabolic reprogramming in polycystic kidney disease reveals coordinated rewiring of bioenergetic pathways. Communications Biology, 1, 194.

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Culturing B-cell Leukemia Cell Lines

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Human cell lines included RS4;11 (pro-B t(4;11), ATCC), SEM (pre-B t(4;11), DSMZ), BV173 (pre-B t(9;22) Ph⁺, DSMZ), and NALM6 (pre-B Ph⁻, ATCC). All cells were cultured in 5% CO₂ at 37°C in RPMI 1640 media (ThermoFisher) with 10% fetal bovine serum (Omega Scientific, Inc), 2 mM glutamine (ThermoFisher), and 1mM sodium pyruvate (ThermoFisher). Murine GFP⁺ 8093 ALL cells (previously described [23 (link),24 (link)]) were cultured in McCoy’s 5A media (Invitrogen) with 10% FBS, 2 mM Glutamax (ThermoFisher), 1 mM sodium pyruvate, 10 ng/ml IL-3 (Peprotech), and 1:1000 BME/2-Mercaptoethanol (Gibco). Human cell lines were authenticated by the University of Arizona Genetics core in November 2016 and tested negative for mycoplasma.

Sea J.L., Orgel E., Chen T., Paszkiewicz R.L., Krall A.S., Oberley M.J., Stiles L, & Mittelman S.D. (2019). Levocarnitine does not impair chemotherapy cytotoxicity against Acute Lymphoblastic Leukemia. Leukemia & lymphoma, 61(2), 420-428.

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Culturing Lenti-X HEK293T and NALM6 Cells

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Lenti-X HEK293T cells (Takara Bio, catalog no. 632180) were maintained in high-glucose Dulbecco’s modified Eagle’s medium with GlutaMAX (Fisher Scientific, catalog no. 10566024), supplemented with 10% FCS, 100 U/ml of penicillin/streptomycin (PenStrep; Fisher Scientific, catalog no. 15140122), 1 mM sodium pyruvate (Fisher Scientific, catalog no. 11360070), 1× minimal essential medium (MEM) nonessential amino acids (Fisher Scientific, catalog no. 11140050), and 10 mM HEPES solution (Sigma-Aldrich, catalog no. H0887–100ML). Cells were passaged every 2 days using Tryple Express (Fisher Scientific, catalog no. 12604013) for dissociation and maintained at <60% confluency.
NALM6 cells were engineered to express NY-ESO-1 peptide in an HLA-A0201 background, recognizable with the 1G4 TCR by the Eyquem laboratory at University of California San Francisco (UCSF) and provided for TCR stimulation coculture experiments. For simplicity, these cells are referred to as NALM6. NALM6 cells were cultured in RPMI (Invitrogen, catalog no. 21870092) supplemented with 10% FCS, 100 U/ml PenStrep (Fisher Scientific, catalog no. 15140122), 1 mM sodium pyruvate (Fisher Scientific, catalog no. 11360070), and 1X MEM nonessential amino acids (Fisher Scientific, catalog no. 11140050), 10 mM HEPES solution (Sigma-Aldrich, catalog no. H0887–100ML), and 2 mM L-glutamine (Lonza Bioscience, catalog no. 17–605E).

Schmidt R., Steinhart Z., Layeghi M., Freimer J.W., Bueno R., Nguyen V.Q., Blaeschke F., Ye C.J, & Marson A. (2022). CRISPR activation and interference screens decode stimulation responses in primary human T cells. Science (New York, N.Y.), 375(6580), eabj4008.

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