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| General Information | |
| Organism | Homo sapiens, human |
| Cell Line Description | HuH-6 is a human hepatoblastoma cell line established in 1974 by Isao Doi and Jiro Sato at Okayama University. It was derived from primary liver tumor tissue obtained from a one-year-old Japanese male infant diagnosed with mixed-type hepatoblastoma. HuH-6 cells exhibit a characteristic polygonal, epithelial-like morphology and grow as a tightly adherent monolayer. In the fields of pediatric oncology, hepatology, and virology, HuH-6 is recognized as the preferred "gold standard" in vitro model for studying human hepatoblastoma. Unlike many other liver-derived cell lines, HuH-6 stably retains the functional characteristics of differentiated hepatocytes, continuously synthesizing and secreting liver-specific proteins such as alpha-fetoprotein (AFP), albumin, prealbumin, and fibrinogen. This cell line is highly valuable for investigating the mechanisms of pediatric liver tumorigenesis, evaluating novel chemotherapeutic agents, and analyzing host-pathogen interactions involving viruses such as hepatitis C virus (HCV) and SARS-CoV-2. |
| Tissue | Liver / Gallbladder |
| Cell Type | Epithelial-like (Hepatoblast) |
| Ethnicity | Asian |
| Disease | Hepatoblastoma |
| Morphology | Epithelial-like; uniform polygonal cells growing in cohesive cobblestone sheets |
| Gender | Male |
| Age | 1 year |
| Growth Mode | Adherent |
| Biosafety Level | 1 (Biosafety classification matches standard laboratory parameters for non-infectious human pediatric tumor cells) |
| Applications | 1. Gold-standard in vitro model for investigating the molecular pathology and signaling networks (specifically the Wnt/β-catenin pathway) of human hepatoblastoma; 2. Standard target line for preclinical high-throughput screening of pediatric-targeted chemotherapeutic agents and tyrosine kinase inhibitors; 3. Modeling the cellular synthesis, processing, and secretion mechanisms of human alpha-fetoprotein (AFP) and albumin; 4. Serving as an alternative host cell line supporting persistent replication of Hepatitis C Virus (HCV) and SARS-CoV-2 clinical isolates; 5. Investigating drug metabolism, enzymatic biotransformation pathways, and mechanisms of multidrug resistance (MDR) in hepatic lineages. |
| Shipped In | Dry ice |
| Storage Temperature | −196°C |
| Characteristics | |
| Tumorigenic | Yes; highly tumorigenic in nude mice, forming progressively expanding tumors that histologically mirror embryonal and fetal hepatoblastoma structures. |
| Transformant | None; immortalized spontaneously from advanced primary pediatric hepatoblastoma tissue. |
| Oncogenic Driver | β-catenin/CTNNB1 mutation. It harbors a classic heterozygous activating missense mutation, p.Gly34Val (c.101G>T), in the CTNNB1 gene. This variant leads to ligand-independent nuclear accumulation of β-catenin, thereby constitutively activating the canonical Wnt transcriptional cascade. |
| Karyotype | Human hyperdiploid/hypertetraploid karyotype with a modal chromosome number of 49 (ranges from 47 to 49). |
| Mycoplasma Test | Negative |
| Culture Conditions and Handling | |
| Subculturing | 1. Remove and discard all spent complete culture medium from the culture vessel. 2. Gently rinse the cell monolayer with sterile, Ca2+/Mg2+-free PBS to neutralize residual serum containing trypsin inhibitors. 3. Add 1.0 to 2.5 mL of pre-warmed 0.2%–0.25% trypsin–0.02%–0.03% (0.53 mM) EDTA solution to completely cover the cell layer. 4. Incubate at 37°C for 3 to 8 minutes; monitor closely under an inverted microscope until the cells round up, loosen, and completely detach. 5. Immediately add an equal volume of complete culture medium to neutralize trypsin activity. 6. Gently pipette the suspension up and down to disperse the cells, centrifuge at ~125 × g for 5 minutes, carefully decant the supernatant, gently resuspend the pellet in fresh complete culture medium, and distribute into new vessels. |
| Thawing Protocol Note | HuH-6 cells are sensitive to the initial alkalinity of the culture medium after thawing. Before adding the thawed cell suspension, the culture vessel containing fresh complete growth medium should be placed in a 5% CO2 incubator for at least 15 minutes to allow the medium to reach and equilibrate within the normal physiological pH range (7.0 to 7.6). |
| Medium Renewal | 2 to 3 times per week |
| Subcultivation Ratio | Split confluent cultures (70-80% density) at a standard ratio of 1:3 to 1:6. |
| Culture Conditions | Atmosphere: Air, 95%; CO2, 5%; Temperature: 37°C |
| Cryopreservation | 90% Complete culture growth medium + 10% DMSO (or 50% Basal medium + 40% FBS + 10% DMSO) |
| Cat.No. | Product Name | Price |
|---|---|---|
| CSC-RO02593 | Cas9 Stable Cell Line - HUH-6 | Inquiry |
| CSC-RR01022 | GFP/Luc Reporter Cell Line - HUH-6 | Inquiry |
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