Sodium pyruvate

Manufactured by R&D Systems

Sodium pyruvate is a chemical compound commonly used in cell culture media. It is a salt of pyruvic acid and provides an energy source for cells in culture. Sodium pyruvate can be utilized by cells as a substrate for various metabolic pathways.

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Lab products found in correlation

Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Recombinant human gm csf, by R&D Systems (1 mentions) Recombinant human il 4, by R&D Systems (1 mentions) T cell isolation kit 2, by Miltenyi Biotec (1 mentions) Ficoll paque, by GE Healthcare (1 mentions) Pen strep, by R&D Systems (1 mentions) Anti cd14 microbeads, by Miltenyi Biotec (1 mentions) Scleraxis, by Abcam (1 mentions) Eya 1, by Santa Cruz Biotechnology (1 mentions) Alpha mem, by Thermo Fisher Scientific (1 mentions) Bm msc, by Lonza (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Penicillin, by Merck Group (1 mentions) Ascorbic acid, by Merck Group (1 mentions) Stempro 34, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by R&D Systems (1 mentions) Interleukin 3 (il 3), by R&D Systems (1 mentions) Stempro 34 nutrient supplement, by Thermo Fisher Scientific (1 mentions)

6 protocols using sodium pyruvate

Monocyte-Derived Dendritic Cell Generation

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Fifty to 70 ml of blood was taken by venous puncture. Peripheral blood mononuclear cells were isolated from heparinized blood by Ficoll-Paque (GE Healthcare) purification. Briefly, blood was diluted 1:1 with sodium chloride and tubes were centrifuged at 400g for 30 minutes with the brake off. Cells were harvested from the interface of the Ficoll layer, and were washed and enumerated. Monocytes were isolated using anti-CD14 microbeads, according to the manufacturer’s instruction (Miltenyi Biotec, San Diego, CA, USA). Cells were cultured in RPMI 1640 with 10% human serum and L-glutamine, pen/strep, non-essential amino acids, sodium pyruvate, 2-ME, 40 ng/ml of recombinant human IL-4 and 40 ng/ml of recombinant human GM-CSF (both from R&D Systems). Cytokines were replenished on days 3 and 6, and cells were used on day 7. For some experiments, CD4⁺ T cells were isolated from the CD14^- fraction using the T cell isolation II kit (Miltenyi Biotec). Cells were then cultured with MoDCs in the presence or absence or RSV. At 48 hours, RNA was extracted and message levels of IFN-γ, IL-5 and IL-13 were determined by qPCR.

Ptaschinski C., Mukherjee S., Moore M.L., Albert M., Helin K., Kunkel S.L, & Lukacs N.W. (2015). RSV-Induced H3K4 Demethylase KDM5B Leads to Regulation of Dendritic Cell-Derived Innate Cytokines and Exacerbates Pathogenesis In Vivo. PLoS Pathogens, 11(6), e1004978.

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Myogenic Differentiation of Stem Cells

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To induce myogenic differentiation, encapsulated GMSCs and hBMMSCs (2×10⁶ cells) in 1 mL alginate microspheres containing the growth factors cocktail were cultured in a medium containing alpha- MEM with 15% FBS, 2 mM L-glutamine, 100 nM Dex, 100 µM ascorbic acid, 2 mM sodium pyruvate (R&D Systems Inc), 100 U/mL penicillin, 100 µg/mL streptomycin, and a cocktail of 10 µM forskolin (FSK), MeBIO, and 10 ng/ml recombinant human bFGF. Cell free RGD-alginate microspheres without any growth factor were used as the control group. Four weeks after induction, the samples were fixed with 4% PFA, and paraffin sections were made. Sections were immunolabeled using antibodies against MF20 (R&D Systems Inc), Myf5, and MyoD (Santa Cruz Biotechnology Inc, Dallas, TX), at 4°C overnight, detected using Alexa fluor conjugated secondary antibody (1:200 dilution; Invitrogen), and counterstained with DAPI.

Ansari S., Chen C., Xu X., Annabi N., Zadeh H.H., Wu B.M., Khademhosseini A., Shi S, & Moshaverinia A. (2016). Muscle tissue engineering using gingival mesenchymal stem cells encapsulated in alginate hydrogels containing multiple growth factors. Annals of biomedical engineering, 44(6), 1908-1920.

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Tenogenic Differentiation of Mesenchymal Stem Cells

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To induce tenogenic differentiation, encapsulated PDLSCs and GMSCs as well as hBMMSCs (2×10⁶ cells in 1 mL of TGF-β3-loaded alginate microspheres) were cultured in a tenogenic medium containing DMEM with 15% FBS, 2 mM L-glutamine, 100 nM Dex, 100 3M ascorbic acid, 2 mM sodium pyruvate (R&D Systems Inc), 100 U/mL penicillin, and 100 μg/mL streptomycin [26 (link)]. Cell-free RGD-coupled alginate microspheres without TGF-β3 were used as the negative control.
Four weeks after induction, the samples were fixed with 4% PFA, and paraffin sections were made. Sections were immunolabeled using antibodies against Tenomodulin (Tnmd), Eya 1, Eya2 (from Santa Cruz Biotechnology, Inc, Dallas, TX), and Scleraxis (Abcam) at 4°C overnight, detected using Alexa fluor conjugated secondary antibody (1:200 dilution; Invitrogen), and counterstained with DAPI.

Moshaverinia A., Xu X., Chen C., Ansari S., Zadeh H.H., Snead M.L, & Shi S. (2014). Application of stem cells derived from the periodontal ligament or gingival tissue sources for tendon tissue regeneration. Biomaterials, 35(9), 2642-2650.

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Isolation and culture of human GMSCs

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Human GMSCs were isolated and cultured according to previously published procedures.^{51 (link),52 (link)} Human bone marrow mesenchymal stem cells (BMMSCs), purchased from Lonza (Gaithersburg, MD).
GMSCs, and hBMMSCs (as a positive control) were separately cultured in a regular culture media containing alpha-MEM (Invitrogen) with 15% FBS, 2 mM L-glutamine (Invitrogen), 100 nM Dex, 100 µM ascorbic acid (Sigma), 2 mM sodium pyruvate (R&D Systems Inc, Minneapolis, MN), 100 U/mL penicillin, and 100 µg/mL streptomycin (Sigma). Passage 4 cells were used in the experiments and hBMMSCs were used as the positive control group.

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Differentiation and Culture of Mast Cells

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Mouse bone marrow-derived mast cells (BMMC) were differentiated from the marrow of tibias and femurs of Wt and Ptch1^+/− littermate mice and cultured for at 6-8 weeks in RPMI 1640 supplemented with 10% FBS, HEPES (1M), penicillin (100 U/mL), streptomycin, 4 mM L-glutamine (100 μg/mL), sodium pyruvate (1 mM), 2-mercaptoethanol (50 μM), and IL-3 (30 ng/mL, R&D Systems). Cultures were analyzed by flow cytometry and BMMC were identified as FcϵRI and Kit double positive cells as described above. Degranulation assays of BMMC were performed as described (26 (link)).
HMC-1.1 and HMC-1.2 cell lines were cultured in Iscove’s Modified Dulbecco’s Medium supplemented with 10% FBS, L-glutamine (2 mM), penicillin (100 U/ml), streptomycin (100 μg/ml). LAD2 cells were cultured in StemPro-34™ supplemented with StemPro-34™ Nutrient Supplement (Gibco), L-glutamine (2 mM), penicillin (100 U/ml), streptomycin (100 μg/ml), and SCF (100 ng/ml, R&D Systems).
Primary human mast cell cultures (hMC) were derived from CD34+ lymphocytapheresis progenitors obtained from healthy volunteers following informed consent under a protocol (NCT00001756) approved by the National Institutes of Health Internal Review Board. Cells were cultured as described (27 (link)) and used between 7–9 weeks of culture when >95% were mast cells identified as KIT and FcεRI double positive cells.

Falduto G.H., Pfeiffer A., Zhang Q., Yin Y., Metcalfe D.D, & Olivera A. (2022). A Critical Function for the Transcription Factors GLI1 and GLI2 in the Proliferation and Survival of Human Mast Cells. Frontiers in Immunology, 13, 841045.

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Differentiation of ND7/23 Sensory Neurons

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DRG neuron‐derived ND7/23 cells (Sigma Aldrich, cat. n. 92090 903) were maintained in 4.5 g/L glucose Dulbecco's Modified Eagle Medium (DMEM, Gibco, cat. n. 52100‐021, UK), 10% fetal calf serum (FCS, Sera Plus, Biotech, cat. n. 3702‐P121812, DE), 1% penicillin/streptomycin (Gibco, cat. n. 15140‐122, UK), and 0.11 g/L sodium pyruvate (Sigma‐Aldrich, cat. n. P5280, JP). The cells were cultured at 37°C and 5% CO₂ and passaged every 4 days.
Passage 8 cells were seeded at 10 000 cells/cm² for the experimental setup. Since neural differentiation of the cell line can establish a long neurite outgrowth which is similar to that observed in primary sensory neurons in culture,³² neural differentiation was induced using 1 mM N6,2′‐O‐Dibutyryladenosine3′,5′‐cyclic monophosphate sodium salt (cAMP, Sigma‐Aldrich, D0260), 10 ng/mL recombinant rat beta‐nerve growth factor (NGF, R&D, 556‐NG‐100), 0.5% FCS in IVD CM or DMEM with 1% penicillin/streptomycin, and 0.11 g/L sodium pyruvate 1 hour after cell seeding and cell attachment.

Ma J., Stefanoska D., Stone L.S., Hildebrand M., van Donkelaar C.C., Zou X., Basoli V., Grad S., Alini M, & Peroglio M. (2020). Hypoxic stress enhances extension and branching of dorsal root ganglion neuronal outgrowth. JOR Spine, 3(2), e1090.

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