HepG2 cells were purchased from the National Collection of Authenticated Cell Cultures, Shanghai, China and STR DNA typing (Dirks and Drexler, 2011 (link)) was used to identify this cell line (results were provided in (results were provided in Supplementary Material S1 ). High-glucose DMEM medium supplemented with 10% fetal bovine serum (FBS) (Universal Biotech Co., Ltd., Shanghai, China) and 1% penicillin/streptomycin (Sangon Biotech Co., Ltd., Shanghai, China) was used to culture the cells in a humidified atmosphere of 95% air and 5% CO2 at 37°C in a cell culture incubator (Memmert GmbH + Co. KG, Schwabach, Germany). Media was replaced at 2-day intervals.
Hepg2 cells
Manufactured by National Collection of Authenticated Cell Cultures
Sourced in China
HepG2 cells are a well-established human hepatocellular carcinoma cell line derived from the liver tissue of a 15-year-old Caucasian male. They are commonly used as an in vitro model for the study of liver biology, metabolism, and toxicology.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (11 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Hepa 1 6 cells, by National Collection of Authenticated Cell Cultures (2 mentions)
Penicillin streptomycin, by Sangon (1 mentions)
Cell culture incubator, by Memmert (1 mentions)
Newborn calf serum, by Sartorius (1 mentions)
Nih3t3 cells, by American Type Culture Collection (1 mentions)
Dmem culture medium, by Thermo Fisher Scientific (1 mentions)
Luciferin potassium salt, by Promega (1 mentions)
Lipofectaminetm 2000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Genview (1 mentions)
Egfp mrna, by TriLink (1 mentions)
Cleancap firefly luciferase mrna, by TriLink (1 mentions)
Tae buffer, by Sangon (1 mentions)
Opti mem 1 reduced serum medium, by Thermo Fisher Scientific (1 mentions)
36 protocols using hepg2 cells
1
Culturing HepG2 Cells for Experiments
2
HepG2 Fatty Acid and Betaine Treatment
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HepG2 cells were purchased from National Collection of Authenticated Cell Cultures (Beijing, China) and cultivated in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum at 5% CO2 and 37°C. When there was an 80% confluence, the cells were seeded into 24-well plates. After 48 h, HepG2 cells were divided into two groups: cells in the OAPA group were cultured with oleic acid (OA, 200 mM) and palmitic acid (PA, 100 mM); cells in the OAPA + Betaine group were cultured with oleic acid, palmitic acid, and betaine (2 mM).
3
Cell Culture Protocols for HepG2 and NIH-3T3
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HepG2 cells were purchased from National Collection of Authenticated Cell Cultures (Shanghai, China). The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), penicillin (100 U/ml), and streptomycin (100 μg/ml) (all from Thermo Fisher), in an atmosphere with 5% CO2. NIH‐3T3 cells were purchased from ATCC (CRL‐1658) (Manassas, VA, USA), and cultured in DMEM with 10% newborn calf serum (Biological Industries, Beit Haemek, Israel).
4
HepG2 cell culture protocol
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HepG2 cells were purchased from the China Center for Type Culture Collection and cultured with Dulbecco’s modified Eagle’s medium (DMEM) culture medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (Gibco, Thermo Fisher Scientific), 100 U/ml of penicillin, and 10 µg/ml of streptomycin in a 37°C incubator with 5% CO2.
5
HepG2 Cell Culture and Mycoplasma Detection
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HepG2 cells were purchased from the China Center for Type Culture Collection and were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 10 µg/ml streptomycin in an incubator containing 5% CO2 at 37°C. The potential presence of mycoplasma in the cell line was detected regularly with a PCR kit (cat. no. C0301S, Beyotime Biotech. Inc., China) followed by the manufacturer's protocol, and no mycoplasma was detected.
6
Formulation and Characterization of mRNA Lipoplexes
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All peptides in this study (Table 1 ) were purchased from ChinaPeptides (Shanghai, China). CleanCap® firefly luciferase mRNA, EGFP mRNA, and cyanine-5 EGFP mRNA were purchased from TriLink Bio Technologies (San Diego, CA, USA). LipofectamineTM 2000 transfection reagent was purchased from Invitrogen (Carlsbad, USA). Luciferin potassium salt was purchased from Promega (Madison, WI, USA). Dulbecco’s modified Eagle’s medium (DMEM), 0.25% Trypsin-EDTA, penicillin/streptomycin antibiotics, and PBS (pH 7.2–7.4) were purchased from Genview (Florida, USA). Fetal Bovine Serum (FBS, South America origin) and Opti-MEM I reduced serum medium were purchased from Gibco (New York, USA). DNA Gel Loading Dye (6 ×) was purchased from Biosharp (Anhui, China). GelRed nucleic acid stain was purchased from US Everbright (California, USA). 1 × Tris-acetate EDTA (TAE) buffer was diluted from 50 × TAE buffer (Sangon Biotech, Shanghai, China) using distilled water. A549 and HepG2 cells were purchased from China Center for Type Culture Collection (CCTCC, Shanghai, China). Other reagents were obtained from Sigma-Aldrich (Saint Louis, MO, USA) of analytical grade or better grade.
7
Culturing HepG-2 Cells in DMEM
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HepG-2 cells were obtained from China Center for Type Culture Collection (CTCC, Wuhan, China), cultured in Dulbecco’s modified Eagle’s medium (DMEM, Hycolne) and supplemented with 10% fetal calf serum (Si Jiqing, Zhejiang Tianhang Biotechnology Corporation, Jinhua, China) with 1% penicillin–streptomycin (Genview, Houston, TX, USA) at 37 °C in a humidified atmosphere of 5% CO2.
8
Cultivation and Palmitate Treatment of HepG2 Cells
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HepG2 cells (China Center for Type Culture Collection, Wuhan, China) were cultured in Dulbecco’s modified Eagle’s medium (Invitrogen, Carlsbad, CA, United States) containing 10% fetal bovine serum (Invitrogen), 100 U/mL penicillin, 100 μg/mL streptomycin, and 1% L-glutamine. Cells were grown at 37 °C in an atmosphere of 5% CO2/95% air in a cell culture flask. The effect of palmitate was examined by addition of this agent to the cells plated in six-well plates at 2 × 105 cells per well.
9
Cultivation of HepG2 Cells with Palmitate
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HepG2 cells, a human hepatocellular carcinoma cell line (China Center for Type Culture Collection, Wuhan, China), were cultivated in Eagle’s medium modified by Dulbecco (DMEM; Invitrogen, Carlsbad, CA, US) supplemented with 100 μg/mL streptomycin, 100 U/mL penicillin, 1% L-glutamine and 10% foetal bovine serum (FBS; Invitrogen). The cells were incubated in cell culture containers at 37°C under a moisturized ambience with 5% CO2 and 95% air until use. The influence of palmitate was investigated by adding this reagent to cells covered in 6-well plates at 2 × 105 cells/well.
10
Gingerol Protects HepG2 Cells from Palmitate-Induced Lipotoxicity
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HepG2 cells were purchased from the China Center for Type Culture Collection (Wuhan, China) and cultured in Minimum Essential Medium (MEM) supplemented with 10% (v/v) fetal bovine serum (FBS), penicillin (100 U/mL) and streptomycin (100 mg/mL) at 37 °C in an atmosphere with 5% CO2. An amount of 10 mM PA/10% fatty acid free bovine serum albumin (BSA) stock solution was prepared according to the protocol as described in a previous study [59 (link)], with a slight modification. When the cell confluence reached about 60%, after 12 h serum starvation, the cells were pretreated with 20 μM or 40 μM 6-gingerol for 24 h. Subsequently, the cells were treated with 200 μM PA plus an indicated concentration 6-gingerol for another 24 h, followed by Nile red staining and the detection of intracellular ROS and TG levels. When using radicicol as an LKB1 destabilizer, the cells were treated with 40 μM 6-gingerol plus 5 μM radicicol in the presence or absence of 200 μM PA for 24 h, followed by indicated experiments.
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