The human HepG2 cell line was purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). HepG2 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) medium (Life Technologies Corporation, Grand Island, NY, USA) containing 10% (v/v) heat-inactivated fetal bovine serum (FBS), 100 units/mL penicillin, 110 mg/L sodium pyruvate, and 100 μg/mL streptomycin (Beyotime, Haimen, China) and maintained in a humidified atmosphere of 95% air and 5% CO2 at 37 °C.
Sodium pyruvate
Manufactured by Beyotime
Sourced in China
Sodium pyruvate is a chemical compound that serves as a source of pyruvate, a key intermediate in cellular metabolism. It is a white crystalline powder that is soluble in water and often used in cell culture media as an energy substrate for cells.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Beyotime (1 mentions)
Streptomycin, by Beyotime (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin and streptomycin, by Beyotime (1 mentions)
Rpmi 1640 medium, by BasalMedia (1 mentions)
Htr 8 svneo, by American Type Culture Collection (1 mentions)
Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions)
Hepes, by STEMCELL (1 mentions)
Anti cd28 antibody, by BioLegend (1 mentions)
Mem nonessential amino acid, by STEMCELL (1 mentions)
Cd8 t cell isolation kit, by BioLegend (1 mentions)
Anti cd3 antibody, by BioLegend (1 mentions)
L glutamine, by STEMCELL (1 mentions)
3 protocols using sodium pyruvate
1
Culturing HepG2 Cells for Biomedical Research
2
Modeling PM2.5 Impact on Trophoblast Cells
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The human trophoblast cell line HTR8/SVneo, originating from human placental trophoblast cells, was purchased from The American Type Culture Collection (CRL-3271, ATCC, USA). The HTR8/SVneo cells we used were identified by STR and tested negative for mycoplasma contamination. HTR8/SVneo was cultured in RPMI-1640 medium (Shanghai BasalMedia Technologies Co., Ltd, China) supplemented with 10% fetal bovine serum (Invitrogen; Thermo Fisher Scientific, Inc, USA), 1% penicillin and streptomycin (Beyotime Institute of Biotechnology, China), and 1% 100 mM sodium pyruvate (Beyotime Institute of Biotechnology, China). Subsequently, the cells were seeded in 6-well plates (Guangzhou Jet Biofiltration Co.,Ltd, China) with 3×105 cells/well, and different concentrations of PM2.5 (50 μg/mL, 100 μg/mL, 200 μg/mL) were added to the culture medium for 24 hr to construct a PM2.5 exposure cell model. N-acetyl-l-cysteine (NAC) (A7250, MillporeSigma, USA) was added to detect the rescue effect on oxidative stress caused by PM2.5 in HTR/SVneo cells.
3
Isolation and Activation of Murine CD8 T Cells
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Isolation of CD8 T cells from the spleens of wild-type C57BL/6 mice was carried out using the CD8 T Cell Isolation Kit (BioLegend, #480035) in accordance with the manufacturer’s instructions. For preactivation, purified CD8 T cells were quantified and seeded onto a flat-bottom 96-well culture plate precoated with 2 µg/mL of anti-CD3 antibody (BioLegend, #100340) and 2 µg/mL of anti-CD28 antibody (BioLegend, #102116). The cells were cultured in a comprehensive medium comprising RPMI 1640, supplemented with 10% FBS, 100 U/mL interleukin-2 (Peprotech, #212–12), 2 mM L-glutamine (STEMCELL Technologies, #07100), 50 µM β-mercaptoethanol (GENOM Biotech, #GNM21985-1), 1 mM sodium pyruvate (Beyotime, C0331), 100 µM MEM non-essential amino acid (STEMCELL Technologies, #07600), and 10 mM HEPES (STEMCELL Technologies, #07200). After 24 hours, the CD8 T cells were co-cultured with macrophages at a 1:1 ratio. Following an additional 48-hour co-culture period, CD8 T cells were subjected to staining for interferon-γ (IFN-γ) and granzyme B (GZMB) to assess their activation status.
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