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| General Information | |
| Organism | Homo sapiens, human |
| Cell Line Description | C3A is a human liver-derived epithelial cell line generated as a clonal derivative of HepG2. The clone was selected for strong contact inhibition, robust production of albumin and alpha-fetoprotein, and the ability to grow under glucose-deficient conditions. C3A retains several differentiated hepatic functions, including synthesis of multiple plasma proteins and measurable nitrogen-metabolizing activity. These properties make it a practical in vitro model for liver biology, hepatic protein production, toxicology, metabolic research, cancer studies, gene delivery, and liver-support system development. |
| Parent Cell Line | HepG2 |
| Tissue | Liver |
| Disease | Hepatoblastoma; historically described as hepatocellular carcinoma |
| Morphology | Epithelial-like; rounded cells may also be observed within the adherent culture |
| Sex | Male |
| Age at Sampling | 15 years |
| Population | White / Caucasian |
| Product Format | Frozen |
| Growth Mode | Adherent |
| Biosafety Level | Biosafety Level 1 |
| Applications | 1. Hepatic physiology and differentiated liver-function research 2. Albumin, alpha-fetoprotein, and plasma-protein production studies 3. Drug response, hepatotoxicity, and chemical safety assessment 4. Glucose utilization, gluconeogenesis, and nitrogen-metabolism research 5. Liver cancer biology and molecular pathway studies 6. Gene expression, transfection, and genome-engineering workflows 7. Bioartificial liver and extracorporeal liver-support research |
| Storage Conditions | Store below −130°C; vapor-phase liquid nitrogen is recommended for long-term preservation |
| Characteristics | |
| Clonal Selection Traits | Selected for strong contact inhibition of growth, high albumin production, high alpha-fetoprotein production, and growth in glucose-deficient medium |
| Tumorigenic | No, under the reported test conditions |
| Hepatic Protein Expression | Produces albumin, alpha-fetoprotein, alpha-2-macroglobulin, alpha-1-antitrypsin, transferrin, alpha-1-antichymotrypsin, haptoglobin, ceruloplasmin, plasminogen, complement C4, fibrinogen, alpha-1-acid glycoprotein, alpha-2-HS-glycoprotein, beta-lipoprotein, and retinol-binding protein |
| Metabolic Characteristics | Exhibits nitrogen-metabolizing activity and oxygen-dependent gluconeogenic activity. Oxygen availability should therefore be considered when designing metabolic experiments. |
| Confluence-Dependent Secretion | As cultures approach confluence, alpha-fetoprotein secretion decreases markedly while albumin secretion increases. Cell density and sampling time should be standardized when comparing secreted-protein measurements. |
| Genetic Profile | A heterozygous NRAS p.Gln61Leu (Q61L; c.182A>T) variant and a TERT promoter C228T variant have been reported. |
| Microsatellite Status | Microsatellite stable (MSS) |
| Mycoplasma Test | Negative |
| Culture Conditions and Handling | |
| Culture Conditions | 37°C in a humidified atmosphere of 95% air and 5% CO₂ |
| Subculturing | 1. Remove and discard the spent culture medium. 2. Briefly rinse the cell layer with 0.25% (w/v) trypsin-0.53 mM EDTA to remove residual serum. 3. Add 2.0-3.0 mL of trypsin-EDTA per 75 cm² flask and monitor under an inverted microscope until the monolayer disperses, usually within 5-15 minutes. Avoid striking or vigorously shaking the vessel during detachment. 4. Add 6.0-8.0 mL of complete medium and gently pipette to prepare an even cell suspension. 5. Transfer the required aliquot to a new culture vessel and incubate at 37°C. |
| Subcultivation Ratio | A split ratio of 1:3 to 1:6 is recommended |
| Medium Renewal | Renew the medium twice per week |
| Thawing and Recovery | Thaw the vial rapidly with gentle agitation in a 37°C water bath, typically within approximately 2 minutes. Transfer the contents to 9.0 mL of complete medium and centrifuge at approximately 125 × g for 5-7 minutes. Resuspend the pellet in fresh complete medium and seed into a suitable culture vessel. Pre-equilibrate the medium to 37°C and normal culture pH before inoculation to reduce recovery stress. |
| Cryopreservation | Complete growth medium supplemented with 5% (v/v) DMSO |
| Handling Notes | Maintain strict aseptic technique and avoid prolonged trypsin exposure or excessive mechanical force. Control confluence carefully because secreted albumin and alpha-fetoprotein change with cell density. For gluconeogenesis studies, standardize oxygenation as well as medium composition, passage history, seeding density, and collection time. |
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