HepG2 cells were obtained from Clontech and cultured at 37 °C and 5% CO2 in complete medium (DMEM; Gibco) containing penicillin/streptomycin (PAA Laboratories) and 10% fetal calf serum (FCS; Anprotec) on cell culture dishes coated with rat-tail collagen (Corning). HepG2 cells were plated on 6-well plates to reach 80–90% confluence on the day of transfection. Cells were transfected with XtremeGeneHP (Roche) following the manufacturer’s instructions. The day after transfection, cells were washed twice with PBS and cultivated in Williams’ E medium (Gibco) containing penicillin/streptomycin, 200 µM L-glutamine (Gibco), 10 mM Hepes (Gibco), and 2% FCS. On day 4 post-transfection, the cell culture supernatant was collected and cells were directly lysed in-well with an “optimized lysis buffer” described previously [41 (link)]. Supernatant and lysates were stored at −20 °C until further use.
Hepg2
Manufactured by Takara Bio
The HepG2 is a human liver cancer cell line commonly used in cell biology research. It is derived from the liver tissue of a 15-year-old Caucasian male with a well-differentiated hepatocellular carcinoma. The HepG2 cell line retains several specialized functions characteristic of normal human hepatocytes, making it a valuable tool for studying liver-related cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
William s e medium, by Thermo Fisher Scientific (1 mentions)
Rat tail collagen, by Corning (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by GE Healthcare (1 mentions)
X tremegene hp, by Roche (1 mentions)
2 protocols using hepg2
1
HepG2 Cell Culture and Transfection
2
Immortalized Hepatocyte-like Cells from Mesenchymal Stem Cells
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The Bmi-1/hTERT-induced human mesenchymal stem cells were previously produced by Sangiamsuntorn et al. [9] , which was established from bone marrow isolated cells. Cells were transformed into the immortalized hepatocyte-like cells using lentiviral transduction with the polycomb group transcription factor Bmi-1, and the human telomerase reverse transcriptase gene (hTERT). The Bmi-1/hTERT-induced immortalized hepatocyte-like cells (imHC) and human hepatocellular carcinoma cells (HepG2) (Clontech, CA) were cultivated in a T-75 tissue culture flask with maintaining media (Dulbecco's modified Eagle's medium: Nutrient mixture F-12; DMEM/F12), supplemented with 10 % fetal bovine serum (FBS), 100 units/mL GlutaMaxTM and 1 % penicillin/streptomycin at 37 ºC in 5 % CO2. Subconfluent imHC at passages 50 -57 were used in all experiments. In the experiments, the cells were seeded on 6-well plates at a density of 106 cells/well in culture media (William's E media plus 10 % FBS, GlutaMaxTM and penicillin/streptomycin. The media was changed daily until the treatment, which used treatment media (William's E serum-free media plus GlutaMaxTM and pen/strep) with C. comosa and an inhibitor (Erythromycin; Eryc) or inducer (Rifampicin; RIF).
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