Sodium pyruvate

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Sodium pyruvate is a chemical compound that functions as an energy source and metabolic intermediate in cell culture media. It is commonly used as a supplement in cell culture applications to support cell growth and metabolism.

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Lab products found in correlation

1 753 protocols using sodium pyruvate

Isolation and Culture of Mammalian Fibroblasts

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Mouse and rat fibroblasts were isolated from ear pieces. Ear pieces were cut in small parts, put in 2–3 wells of a 6-well plate (Nunc) containing 50 µl fibroblast culture media (RPMI 1640 (Millipore, F1215), 20% (v/v) FCS (Gibco by life technologies, 10270), 1% (v/v) Pen/Strep (Millipore, A2213), 10 mM HEPES (Gibco by life technologies, 15630-056), 1 mM Sodium pyruvate (Millipore, L0473), 1x NEAA (Millipore, K0293), 1x Glutamax-I (Gibco by life technologies, 35050-038)), covered with 2 coverslips and flooded with 3 ml fibroblasts culture media. The media was changed every 3–4 days. Growing fibroblasts were split using Accumax solution (Sigma, A7089) after the cells become confluent. For all experiments fibroblasts in the passage 3 to 7 were used.
All human fibroblasts from Parkinson patients and healthy control individuals are from the Blood- and Tissue-library Hertie Institute for Clinical Brain Research Tuebingen (Ethics-Application 287/2004V). Human fibroblasts are cultured in RPMI media (Millipore, F1215) containing 10% (v/v) FCS (Gibco by life technologies, 10270), 1 mM Sodium pyruvate (Millipore, L0473) and 1x Glutamax-I (Gibco by life technologies, 35050-038).

Funk N., Munz M., Ott T., Brockmann K., Wenninger-Weinzierl A., Kühn R., Vogt-Weisenhorn D., Giesert F., Wurst W., Gasser T, & Biskup S. (2019). The Parkinson’s disease-linked Leucine-rich repeat kinase 2 (LRRK2) is required for insulin-stimulated translocation of GLUT4. Scientific Reports, 9, 4515.

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Metabolic Tolerance Tests in Mice

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For the lactate or pyruvate tolerance tests, fed mice were injected IP with lactic acid solution (0.5 g/kg body weight) (MilliporeSigma) or sodium pyruvate (1.0 g/kg body weight) (MilliporeSigma) formulated in PBS. For the gluconeogenesis assay, the mice were fasted for 16 hours before being administered IP with sodium lactate (2.5 g/kg body weight), sodium pyruvate (2.5 g/kg body weight), alanine (2.5 g/kg body weight) (MilliporeSigma), or l-glutamine (2.5 g/kg body weight) (MilliporeSigma) formulated in PBS. For glucose tolerance tests, Bsg^+/+ and Bsg^–/– mice fasted overnight were administered glucose (1 g/kg body weight), either by IP injection or by oral gavage. Blood glucose and lactate levels were measured using a glucometer (Johnson & Johnson K.K.) and a lactate test meter (ARKRAY), respectively. Serum pyruvate values were determined using a pyruvate colorimetric assay kit (catalog K609; BioVision, Inc.) according to the manufacturer’s protocol. The x axis was used as the baseline of the AUCs in our calculations.

Ryuge A., Kosugi T., Maeda K., Banno R., Gou Y., Zaitsu K., Ito T., Sato Y., Hirayama A., Tsubota S., Honda T., Nakajima K., Ozaki T., Kondoh K., Takahashi K., Kato N., Ishimoto T., Soga T., Nakagawa T., Koike T., Arima H., Yuzawa Y., Minokoshi Y., Maruyama S, & Kadomatsu K. (2021). Basigin deficiency prevents anaplerosis and ameliorates insulin resistance and hepatosteatosis. JCI Insight, 6(20), e142464.

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Hydrogen Peroxide Scavenging Assay

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H₂O₂ scavenging assay, based on transition metal chelation, was modified from Hazra et al. [50 (link)] and Jiang et al. [51 (link)] with microplate reader in 96-well format with four technical replicates on each plate. In the assay the formation of ferric-xylenol orange complex indicates the ability of the sample to scavenge H₂O₂ and prevent the oxidation of Fe(II) to Fe(III). Each sample was measured using the extract (1:10) without dilutions and the result is expressed as inhibition percentage (%) of Fe(II) oxidation to Fe(III).
In the assay the sample is mixed with an aliquot of 2 mM H₂O₂, 2.56 mM ammonium iron (II) sulphate·6H₂O and 111 µM xylenol orange disodium salt and after 30 min incubation, the absorbance of ferric-xylenol orange complex at 560 nm was measured.) Sodium pyruvate (Sigma-Aldrich Chemie GmbH, St. Louis, MO, USA) was used as a reference compound. Reagents: H₂O₂ (Merck KGaA, Darmstadt, Germany), ammonium iron (II) sulphate·6H₂O (BDH Prolabo, Dubai, UAE), xylenol orange disodium salt (Sigma-Aldrich Chemie GmbH, St. Louis, MO, USA), Sodium pyruvate (Sigma-Aldrich Chemie GmbH, St. Louis, MO, USA).

Korkalo P., Korpinen R., Beuker E., Sarjala T., Hellström J., Kaseva J., Lassi U, & Jyske T. (2020). Clonal Variation in the Bark Chemical Properties of Hybrid Aspen: Potential for Added Value Chemicals. Molecules, 25(19), 4403.

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Cell Culture Conditions for BS153 and DKMG

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BS153vIII+/-and DKMGvIII+/-sublines have recently been described in detail. 21 BS153 sublines were cultured in DMEM (Sigma-Aldrich) supplemented with 10% heat inactivated FCS, 2 mM L-glutamine and 1 mM sodium pyruvate (Sigma-Aldrich); DKMG cells were cultured in RPMI (10% heat inactivated FCS, 2 mM L-glutamine, 1 mM sodium pyruvate). All cells were grown at 37 °C, 5% CO 2 and 100% humidification. All cells were identified by a short tandem repeat multiplex assay (Applied Biosystems).

Boyd P.S., Struve N., Bach M., Eberle J.P., Gote M., Schock F., Cremer C., Kriegs M, & Hausmann M. (2016). Clustered localization of EGFRvIII in glioblastoma cells as detected by high precision localization microscopy. Nanoscale, 8(48).

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Cultivation of Cancer Cell Lines

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The epithelial human glioblastoma cell line U87 MG (ATCC HTB-14, MGMT^-) and the human hepatocellular carcinoma cell line HepG2 (ATCC HB-8065) were grown in complete growth medium, consisting of Dulbecco’s Eagle’s Minimum Essential Medium (DMEM) (Sigma-Aldrich, Steinheim Germany) supplemented with 10% v/v heat inactivated fetal calf serum (FCS) (Sigma-Aldrich), 1% antibiotics (10,000 IU/mL penicillin, 10 mg/mL streptomycin, Sigma-Aldrich), l-glutamine (Sigma-Aldrich), MEM non-essential amino acid solution 100× (Sigma-Aldrich) and sodium pyruvate (Sigma-Aldrich).The epithelial human cervix carcinoma cell line HeLa (ATCC CCL-2) was grown in complete growth medium, consisting of RPMI-1640 (Sigma-Aldrich, Germany) supplemented with 10% v/v heat inactivated FCS (Sigma-Aldrich), 1% antibiotics (10,000 IU/mL penicillin, 10 mg/mL streptomycin, Sigma-Aldrich), l-glutamine (Sigma-Aldrich) and sodium pyruvate (Sigma-Aldrich). After reaching confluency, adherent growing tumor cells were trypsinized for 2 min at 37 °C in trypsin ethylene diamine-tetra-acetic acid (EDTA) (Sigma-Aldrich). Single cell suspensions with different cell counts were seeded in 15 mL supplemented medium in T-75 ventilated culture flasks. Tumor cells were routinely checked for mycoplasma contamination.

Fellinger H., Stangl S., Hernandez Schnelzer A., Schwab M., Di Genio T., Pieper M., Werner C., Shevtsov M., Haller B, & Multhoff G. (2020). Time- and Dose-Dependent Effects of Ionizing Irradiation on the Membrane Expression of Hsp70 on Glioma Cells. Cells, 9(4), 912.

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Casein-Casein-Yeast Medium for PVL Production

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The casein–casein–yeast (CCY) medium was prepared to contain 30 g/L yeast extract (Oxoid, Basingstoke, UK), 20 g/L casamino acids (VWR Amresco, Cleveland, OH, USA), 2.48 g/L Na₂HPO₄, 0.41 g/L KH₂PO₄, 20 mg/L MgSO₄·7H₂O, 7.5 mg/L MnSO₄·H₂O, 6.4 mg/L citric acids, and 6.4 mg/L FeSO₄·7H₂O (Sigma, Darmstadt, Germany) (pH 7.3 ± 0.2 at 25 °C). Then, a 100 mL sodium pyruvate solution was prepared by dissolving 23.2 g of sodium pyruvate (Sigma) in 100 mL deionised water, sterilised by 0.22 µm filtration, and added to 900 mL autoclaved CCY medium at 121 °C for 15 min [32 ]. S. aureus test strains, M1, YF1B-b, and S. aureus control strain HT480, were grown on a Tryptone Soy Agar (CM131, Oxoid) at 37 °C by overnight incubation. Bacterial suspensions were prepared in 0.9% (w/v) NaCl (Sigma) and adjusted to McFarland 0.5 using a densitometer (Den-1B, Biosan, Riga, Latvia). Each bacterial suspension (500 µL) was inoculated to a 50 mL CCY medium and incubated at 37 °C for 24 h by shaking at 120 rpm to promote PVL production.

Yildirim F., Sudagidan M., Aydin A., Akyazi I., Bayrakal G.M., Yavuz O, & Gurel A. (2022). In Vivo Pathogenicity of Methicillin-Susceptible Staphylococcus aureus Strains Carrying Panton–Valentine Leukocidin Gene. Life, 12(12), 2126.

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Modulating Oxidative Phosphorylation in Cells

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For inhibition of oxidative phosphorylation, explants were cultured for 24 hours in DM with 35 mM glucose. Immediately prior to imaging, media was exchanged with DM with PBS vehicle, Oligomycin (Sigma), or Bongkrekic Acid (Enzo Life Sciences). We found a strain difference in response to treatment to BA. CD1 cells were treated with 0.5, 2.5, 5, and 50 μM BA, while Ant1^+/+ and Ant1^−/− were treated with 0.5 fM BA. For glucose deprivation and inhibition experiments, explants were cultured for 24 hours in glucose free DMEM (Invitrogen) plus N2 supplement (Gibco) with or without 10 mM sodium pyruvate (Sigma) or 5 mM Galactose (Sigma), +/− 2.5 μM BA. For treatment with 2-DG, explants were cultured for 24 hours with 5 mM glucose DM supplemented with 500 μM 2-DG (Sigma), with or without 10 mM sodium pyruvate or 5 mM Galactose +/− 0.5 μM BA.

Lin-Hendel E.G., McManus M.J., Wallace D.C., Anderson S.A, & Golden J.A. (2016). Differential mitochondrial requirements for radially and non-radially migrating cortical neurons: Implications for mitochondrial disorders. Cell reports, 15(2), 229-237.

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Chondrogenic and Hypertrophic Differentiation of BMSCs

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For chondrogenic and hypertrophic differentiation,¹⁴ (link) 1 × 10⁶ BMSCs were placed in polypropylene tubes, centrifuge at a speed of 500 g for 20 min. Then remove the medium and add 500 μL of low-glucose DMEM supplemented with 10⁻⁷ M dexamethasone (Sigma–Aldrich), 1%(v/v) ITS (Sigma–Aldrich), 50 μM ascorbate-2-phosphate (Sigma–Aldrich), 1 mM sodium pyruvate, 50 μg/ml proline (Sigma–Aldrich) and 20 ng/ml TGFβ3. Change the medium three times a week and culture for 14d. Then, the serum-free chondrogenic medium was replaced with hypertrophic medium supplemented with 50 nM thyroxine (Sigma–Aldrich), 10⁻⁸ M dexamethasone, 250 μM ascorbate-2-phosphate, 1 mM sodium pyruvate and 50 μg/ml proline. Change the medium three times a week and culture for another 14d.

Bai Y., Gong X., Dong R., Cao Z., Dou C., Liu C., Li J., Kang F., Dai J., Zhao C., Tian Z., Tan J., Dai Q, & Dong S. (2020). Long non-coding RNA HCAR promotes endochondral bone repair by upregulating VEGF and MMP13 in hypertrophic chondrocyte through sponging miR-15b-5p. Genes & Diseases, 9(2), 456-465.

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Propagation and Maintenance of H1299-E3 and VeroE6 Cell Lines

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The H1299-E3 (H1299-ACE2, clone E3, H1299 originally from ATCC as CRL-5803) cell line was derived from H1299 as described in our previous work (Cele et al. 2021a (link), 2021b (link)). The H1299-E3 cells were propagated in growth medium consisting of complete Roswell Park Memorial Institute (RPMI) 1640 medium with 10 per cent fetal bovine serum (Hyclone) containing 10 mM of hydroxyethylpiperazine ethanesulfonic acid (HEPES), 1 mM sodium pyruvate, 2 mM l-glutamine and 0.1 mM nonessential amino acids (all Sigma-Aldrich). Cells were passaged every second day. For virus isolation, Vero E6 cells (originally ATCC CRL-1586, obtained from Cellonex in South Africa) were propagated in complete growth medium consisting of Dulbecco’s Modified Eagle Medium (DMEM) with 10 per cent fetal bovine serum (Hyclone) containing 10 mM of HEPES, 1 mM sodium pyruvate, 2 mM l-glutamine and 0.1 mM nonessential amino acids (all Sigma-Aldrich). Vero E6 cells were passaged every 3–4 days. The VeroE6 cells expressing TMPRSS2 and ACE2 (VeroE6-TMPRSS2), originally BEI Resources, NR-54,970 were used for virus re-expansion of the virus from the day 6 swab and fusion assay. The Vero-TMPRSS2 cell line was propagated in the same way as VeroE6 cells.

Lustig G., Ganga Y., Rodel H.E., Tegally H., Khairallah A., Jackson L., Cele S., Khan K., Jule Z., Reedoy K., Karim F., Bernstein M., Ndung’u T., Moosa M.Y., Archary D., de Oliveira T., Lessells R., Neher R.A., Abdool Karim S.S, & Sigal A. (2023). SARS-CoV-2 infection in immunosuppression evolves sub-lineages which independently accumulate neutralization escape mutations. Virus Evolution, 10(1), vead075.

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Culturing Oral Cancer Cell Lines

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The SAS and OECM-1 human oral cancer cell lines were purchased from ATCC (ATCC: American Type Culture Collection, Manassas, VA, USA). SAS cells were cultured in Dulbecco's modified Eagle's medium-F12 supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, 1.5 g/L sodium bicarbonate, 25 mM HEPES and 1 mM sodium pyruvate (Sigma, St. Louis, Mo, USA). OECM-1 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with 10% FBS, 1% penicillin/streptomycin, 1.5 g/L sodium bicarbonate, 25 mM HEPES and 1 mM sodium pyruvate (Sigma, St. Louis, Mo, USA). The cells culture weas maintained at 37°C in a humidified atmosphere of 5% CO₂.

Hsieh M.J., Lin C.W., Chiou H.L., Yang S.F, & Chen M.K. (2015). Dehydroandrographolide, an iNOS inhibitor, extracted from from Andrographis paniculata (Burm.f.) Nees, induces autophagy in human oral cancer cells. Oncotarget, 6(31), 30831-30849.

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