HuH-6 Cells
Product number:
305092
General information
Description | HuH-6 cells, short for Human Hepatoma 6 cells, are a permanent cell line established from hepatoma tissue obtained from a 57-year-old Japanese male in 1982. These cells have become a valuable tool in biological research, providing a convenient substitute for primary hepatocytes. The primary application of HuH-6 cells lies in studying hepatitis C virus (HCV) infection and hepatoma. Researchers often use these cells as a cell culture model to investigate the mechanisms of HCV infection and explore potential treatments for hepatoma. One crucial area of study involving HuH-6 cells is understanding drug metabolism and pharmacokinetics. These cells are commonly used to assess the potential for drug-drug interactions (DDIs) by analyzing the gene expression of various CYP450 enzymes. Regulatory agencies require an evaluation of DDI potential in hepatocytes for specific enzymes like CYP3A4, CYP1A2, and CYP2B6. HuH-6 cells, known as Cryopreserved Human Hepatocytes, Induction qualified (HUCPI), have been characterized to exhibit enzyme induction levels recommended by the FDA for evaluating DDIs. Another significant application of HuH-6 cells is the study of transporter-mediated efflux inhibition. Plating the hepatocytes onto collagen and overlaying them with a basement membrane extract allows the formation of polarized cell surface structures and the re-expression of bile efflux transporters. This enables researchers to investigate the effects of drug inhibition on bile efflux transport, which can range from drug-drug interactions to liver toxicity caused by the accumulation of intracellular bile acids. Hepatocytes pre-characterized for transporter function are well-suited for these studies. Additionally, HuH-6 cells play a crucial role in the pharmaceutical discovery of drugs with a slower metabolism. By measuring the basic metabolic properties of drugs that are metabolized at a slower rate, researchers can develop medications that remain effective for more extended periods. Plated hepatocytes, like HuH-6 cells, maintain their metabolic activity for an extended duration, making them ideal for assays lasting four hours or longer. Hepatocytes prequalified for low clearance are especially valuable for these studies, although Induction or Transporter qualified hepatocytes (HUCPI) can also be used. HuH-6 cells are a valuable cell line derived from hepatoma tissue. They are commonly used in biological research to study hepatitis C virus infection, hepatoma, drug metabolism and pharmacokinetics, drug-drug interactions, transporter-mediated efflux inhibition, and the discovery of drugs with a slower metabolism. These cells provide researchers with a convenient and reliable model to investigate various aspects of liver-related biology and drug development. |
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Organism | Human |
Tissue | Liver |
Disease | Hepatoblastoma |
Synonyms | HUH-6, HuH 6, HuH6, HUH6, Huh6 |
Characteristics
Age | 1 year |
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Gender | Male |
Ethnicity | Asian |
Morphology | Epithelial |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | HuH-6 (Cytion catalog number 305092) |
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Biosafety level | 1 |
Expression / Mutation
Handling
Culture Medium | DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | 1:2 to 1:4 |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures | HuH-6 cells are shipped in a deep-frozen state on dry ice. Upon receipt, confirm that the vial remains frozen. For storage, place the cryovial immediately at temperatures below -150 degrees. If you plan to culture the cells immediately, swiftly thaw the vial by shaking it in a 37 degrees water bath with clean water and an antimicrobial agent for 40-60 seconds. Remove the vial once a small ice clump persists, ensuring it remains cold. Proceed with all subsequent steps under aseptic conditions. In a sterile flow hood, disinfect the cryovial with 70% ethanol. Then, gently open the vial and transfer the cell suspension into a 15 ml centrifuge tube pre-filled with 8 ml of room temperature culture medium. Gently mix the cells. For cell separation, centrifuge at 300 x g for 3 minutes and dispose of the supernatant. Skipping centrifugation is optional, although any residual freezing medium should be removed after 24 hours. Resuspend the pellet gently in 10 ml of fresh culture medium and divide between two T25 culture flasks. Follow the subculture protocol for subsequent steps. |
Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is rigorously excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
Amelogenin: x,y
CSF1PO: 10,12
D13S317: 8,12
D16S539: 10,11
D5S818: 8,11
D7S820: 11,12
TH01: 7,8
TPOX: 8
vWA: 14,17
D3S1358: 14,17
D21S11: 29,30
D18S51: 13,21
Penta E: 11
Penta D: 9,13
D8S1179: 10,11
FGA: 19,24
D6S1043: 13,18
D2S1338: 18
D12S391: 18,20
D19S433: 12,12.2
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