We collected 40 pairs of liver tumors and adjacent tissues from patients treated at The First Affiliated Hospital of Chongqing Medical University. Patients were excluded if they receive chemotherapy or radiation before surgery. Our study was approved by the Ethics Committee of Chongqing Medical University and that this was conducted in accordance with the Declaration of Helsinki. We obtained written informed consent from each patient after providing information about the study, and patients signed the informed consent forms. The normal hepatocyte cell line (HL-7702) and human HCC lines (SMMC-7721, Hep-3B, Hep-G2, SMMC-7402, and Huh-7) were purchased from the Institute of Chinese Academy of Sciences. SMMC-7721, Hep-3B, Hep-G2, SMMC-7402 and Huh-7 cell lines were cultured in DMEM (Corning Incorporated, Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Thermo Scientific, Waltham, MA, USA). The HL-7702 cell line was cultured in RPMI 1640 medium (Corning Incorporated, Corning, NY, USA) supplemented with 10% FBS. All cells were cultured in an incubator with an atmosphere of 95% O2 and 5% CO2 at 37°C.
Hep3b
Manufactured by Corning
Sourced in United States
The Hep3B is a cell line derived from human hepatocellular carcinoma, used for in vitro studies. The Hep3B cell line retains several characteristics of normal hepatocytes and is a valuable tool for research related to liver function and disease.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Corning (2 mentions)
Hepg2, by Corning (1 mentions)
Smmc 7721, by Corning (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
High glucose dmem, by Corning (1 mentions)
Hcclm3, by Corning (1 mentions)
Hep3b, by iCell Bioscience (1 mentions)
Rpmi 1640, by Corning (1 mentions)
Sorafenib, by Merck Group (1 mentions)
Bobcat339, by MedKoo Biosciences (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Hep3b, by American Type Culture Collection (1 mentions)
Dmem f12 medium, by Corning (1 mentions)
Mem medium, by Corning (1 mentions)
Hep3b, by European Collection of Authenticated Cell Cultures (1 mentions)
Cs fbs, by Merck Group (1 mentions)
Hek 293, by European Collection of Authenticated Cell Cultures (1 mentions)
Mcf 7, by Corning (1 mentions)
Hek293, by Corning (1 mentions)
5 protocols using hep3b
1
Profiling Liver Tumor Samples
2
Culturing Liver Cancer Cell Lines
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Both the liver cancer cell lines Hep3B and HCCLM3 (iCell Bioscience Inc, Shanghai, China) were used in this study. Hep3B cells were grown in high‐glucose DMEM (Corning, China) medium, while HCCLM3 cells were cultured in RPMI‐1640 (Corning, China) medium. Both types of cells were cultured in a medium supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin at a temperature of 37°C and 5% CO2.
3
Cell Line Authentication and Epigenetic Modulation
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The Hep3b and Huh7 cell lines were obtained from American Type Culture Collection with no further authentication or testing for mycoplasma. The cell lines were grown in MEM (Hep3b, CORNING #10-010-CV) or DMEM (Huh7, CORNING #10-013-CV), supplemented with 10% FBS (Gibco by Life Technologies™, #16140-071) and Antibiotic-Antimycotic (Gibco by Life Technologies™, #15,240,062) at 37 °C under 5% CO2. No cell line used in this paper is listed in the database of commonly misidentified cell lines maintained by ICLAC (International Cell Line Authentication Committee). The shRNAs against DNMT3a (V2LHS_202509, V2LHS_74666, V2LHS_74668), TET2 (V2LHS_227656, V2LHS_266084) and the negative control (pGIPZ) vectors were obtained from BMGC RNAi (University of Minnesota). TET2 inhibitor Bobcat339 (#408,006) was obtained from MedKoo Biosciences, and Sorafenib (#SML2653) was obtained from Sigma-Aldrich. Solutions of Bobcat339 were prepared in dimethyl sulfoxide (DMSO) at 100 mM, while Solutions of Sorafenib were prepared in DMSO at 10 mM.
4
Hypoxic Conditioning of Human MSC
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HCC cell lines 7402 and Hep3b were purchased from the Shanghai Institutes for Biological Sciences. HCC cell line 7402 was cultured in RPMI 1640 medium (Corning Inc., Corning, NY, USA), and Hep3b was cultured in MEM medium (Corning Inc.). Both mediums were supplemented with 10% fetal calf serum (FBS), 100 U/ml penicillin and 100 g/ml streptomycin. Cells were maintained in a humidified incubator with 5% CO2 at 37 °C. Human umbilical MSC were purchased from ScienCell (San Diego, California, USA) and cultured in DMEM/F-12 medium (Corning Inc.). MSC at passage 3–10 were used for experiments. When MSC grew to about 80% confluence, they were switched to serum-free medium and cultured for 48 h at 37 °C under hypoxic (1.5% O2) (hypo-MSC) or normoxic (21.0% O2) conditions. The condition medium (CM) was harvested, purified by centrifugation and frozen at − 80 °C for further experiment.
5
Cell Culture Protocols for Hep3B, HEK293, and MCF-7
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All cell culture media and supplements except charcoal-stripped foetal bovine serum, CsFBS (Sigma-Aldrich) were obtained from Life Technologies (Paisley, U.K.). Hep3B (ECACC 86062703) and HEK293 (ECACC 85120602) were obtained from the European Collection of Authenticated Cell Cultures (ECACC) through Sigma-Aldrich. MCF-7 cells were kindly provided by Matilda Katan and Ivan Gout (University College London, U.K.). All cell lines (passage numbers <20) were routinely cultured in DMEM (with 25 mM HEPES, GlutaMAX-1 and 4.5 g/l glucose), supplemented with 10% FBS and antibiotics and antimycotics. For transfections, cells were passaged in phenol red-free DMEM supplemented with 10% CsFBS without antibiotics and antimycotics, and seeded at a confluence of 80–90% in 24-multiwell uncoated plates (Hep3B and MCF-7) or on BioCoat poly D-lysine plates (HEK293); both sets of plates were obtained from Corning (Appleton Woods, U.K.).
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