HepG2 cells
Description
This cell line has been licensed to Revivocell. The sale of these cells requires the completion of a Material Transfer Agreement (MTA). Please contact us for further information.
HepG2 cells are a hepatoblastoma cell line widely used in biological science research. Initially mistaken as hepatocellular carcinoma, they offer valuable insights into various aspects of liver-related studies. These cells were first isolated in 1975 and later recognized as hepatoblastoma rather than hepatocellular carcinoma, resolving years of confusion. Cytologically, HepG2 cells have an average diameter of 10-20 micrometres, large nuclei with 3-7 nucleoli, and low mitochondrial content. They exhibit specific chromosomal abnormalities associated with hepatoblastomas, such as translocations and trisomies. Furthermore, these cells carry a TERT promoter mutation (C228T) and maintain wild-type TP53, essential in cancer development. HepG2 cells differ from normal hepatocytes regarding cytochrome P450 (CYP) enzyme expression. They have a weak or absent expression of several CYP enzymes involved in xenobiotic metabolism in the liver. This distinction should be considered when using HepG2 cells to predict xenobiotic metabolism and elimination. Despite these limitations, HepG2 cells have been utilized in studying drug metabolism and toxicity testing. They exhibit similarities in the expression of drug metabolism/transport proteins in hepatocellular carcinoma and hepatoblastoma cells. Efforts are underway to enhance the expression of cytochromes in HepG2 cells, making them a more reliable model for hepatocyte research. Three-dimensional spheroid cultures are also being explored to create a more physiologically relevant system, as these models exhibit higher metabolic activity than traditional 2D cell cultures.
Product Sheet Safety Data Sheet
Organism
Human
Tissue
Liver
Applications
This cell line is an optimal choice for transfection. Further, the HepG2 cells offer an array of applications, ranging from 3D cell culture and cancer research to high-throughput screening and toxicology.
Characteristics
| Age | 15 years |
|---|---|
| Gender | Male |
| Ethnicity | Caucasian |
| Morphology | Epithelial-like |
| Growth properties | Adherent |
Biosafety level
1
Expression / Mutation
| Receptors expressed | insulin, insulin-like growth factor II (IGF II) |
|---|---|
| Protein expression | p53 positive |
| Tumorigenic | no |
| Products | Albumin, alpha-fetoprotein (alpha fetoprotein), alpha1 acid glycoprotein (alpha-1 acid glycoprotein), alpha1 antitrypsin (alpha-1-antitrypsin), alpha1 antichymotrypsin, (alpha-1-antichymotrypsin), alpha2 HS glycoprotein (alpha-2-HS- glycoprotein), alpha2 macroglobulin (alpha-2-macroglobulin), beta lipoprotein (beta-lipoprotein), ceruloplasmin, C4 and C3 activator, fibrinogen, haptoglobin, plasminogen, retinol binding protein (retinolbinding protein), transferrin |
| Karyotype | modal number = 55 (range = 50 to 60), has a rearranged chromosome 1 |
Handling
| Culture Medium | Ham's F12 |
|---|---|
| Medium supplements | 10% FBS, w: 1.0 mM stable Glutamine, w: 1.0 mM Sodium pyruvate, w: 1.1 g/L NaHCO3 |
| Passaging solution | Accutase |
| Doubling time | 48 hours |
| Subculturing | Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add Accutase (1-2ml per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 5 min at 300xg, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium. |
| Split ratio | A ratio of 1:4 to 1:6 is recommended |
| Seeding density | 2 to 3 x 10^4 cells/cm^2 during routine culture |
| Fluid renewal | 2 to 3 times per week |
| Freezing recovery | Start culture using the complete contents of the cryovial in 2xT25 cell culture flasks. The cells will recover within 48 to 72 hours. |
| Freeze medium | CM-1 (CLS catalog number 800100) or CM-ACF (CLS catalog number 806100) |
| Handling of cryopreserved cultures | The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still frozen. If immediate culturing is not intended, the cryovial must be stored below -150 degree Celsius after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 37 degree Celsius water bath within 40-60 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath. A small ice clump should still remain and the vial should still be cold. From now on, all operations should be carried out under aseptic conditions. Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300 x g for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this case the remains of the freeze medium have to be removed 24 hours later. Resuspend the cells carefully in 10 ml fresh cell culture medium and transfer them into two T25 cell culture flasks. All further steps are described in the subculture section. |
| Handling of proliferating cultures | One or two cell culture flasks come filled with cell culture medium. Collect the entire medium in 1 or 2 x 50 ml centrifuge tubes, respectively. Carefully add 5 ml of cell culture medium to each T25 cell culture flask. Control the cell morphology and confluency under the microscope. Incubate at 37 degree Celsius for a minimum of 24 hours. Spin down the collected medium at 300 x g for 3 minutes to collect the cells which may have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell culture medium and transfer to a T25 cell culture flask. Incubate at 37 degree Celsius for a minimum of 24 hours. |
Quality control / Genetic profile / HLA
| Sterility | Mycoplasma contamination was excluded through PCR-based and luminescence-based mycoplasma assays. Bacterial or fungal contaminations are detected through daily visual cell monitoring. |
|---|---|
| STR profile | Amelogenin: x,Y CSF1PO: 10,11 D13S317: 9,13 D16S539: 12,13 D5S818: 11,13 D7S820: 10 TH01: 9 TPOX: 8,9 vWA: 17 D3S1358: 15,16 D21S11: 29,31 D18S51: 13,14 D8S1179: 15,16,17 FGA: 22,25 D2S1338: 19,20 D19S433: 15.2 |
| HLA alleles | A*: 02:01:01, 24:02:01 B*: 35:14:01, 51:08:01 C*: 04:01:01, 16:02:01 DRB1*: 13:02:01, 16:02:01 DQA1*: 01:02:01, 05:05:01 DQB1*: 03:01, 06:04 DPB1*: 02:01:02, 04:02:01 E: 01:01:01 |