Product Details

• Catalog Number: T0054
• Unit Size: 1×10⁶ cells / 1.0 mL
• Species: Human (Homo sapiens)
• Tissue of Origin: Liver
• Cell Type: Immortalized Hepatocytes
• Growth Properties: Adherent, epithelial morphology
• Immortalization Method: Lentiviral transduction with HPV E6/E7, hTERT & MycT58A
• Expression Markers: AAT1, CK8, CK18, CYP1A1 (constitutive), CYP1A2 (inducible), CYP2C9 (inducible)
• Biosafety Level: BSL-2
• Storage: Below -130°C (liquid nitrogen vapor phase)
• Shipping: Shipped on dry ice
• Format: Cryopreserved frozen cells

Overview

The Immortalized Human Hepatocytes (HPV E6/E7, hTERT & MycT58A) provide a reproducible and long-term in vitro model for investigating hepatic metabolism, liver function, drug toxicity, and disease mechanisms. Their stable genetic background and optimized growth conditions make them an essential tool for liver research.

Key Features and Benefits

• Long-Term Stability: Maintains hepatocyte characteristics over multiple passages without senescence.
• Reproducibility: Ensures experimental consistency with a stable genetic profile.
• Ideal for Liver Research: Suitable for studies on hepatocyte metabolism, proliferation, and toxicity screening.
• Optimized Growth Conditions: Adapted to specialized media and extracellular matrices for enhanced viability.
• Adherent Growth Properties: Facilitates easy handling and maintenance in culture.

Culture & Handling Guidelines

Recommended Culture Conditions

• Coating: Use PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) for optimal adhesion.
• Growth Medium: PriGrow IX (TM019) supplemented with: 
  • 10% Fetal Bovine Serum (FBS)
  • 1% Penicillin/Streptomycin (G255)
• Incubation Conditions: 37°C, 5% CO₂
• Seeding Density: 30,000 cells/cm²
• Doubling Time: ~30 – 35 hours

Thawing Protocol

1. Quickly thaw cells in a 37°C water bath for no more than 2 minutes. Keep the vial cap above the water level to avoid contamination.
2. Decontaminate the vial by spraying with 70% ethanol and transfer to a biological safety cabinet.
3. Add thawed cells to a sterile 15mL conical tube containing 5mL pre-warmed complete growth medium.
4. Centrifuge at 125xg for 5–7 minutes.
5. Aspirate the supernatant, gently resuspend the cell pellet in fresh growth medium, and seed into a pre-coated T25 flask.
6. Incubate under recommended conditions and monitor for cell recovery.

Subculture Guidelines

1. Aspirate the culture medium and rinse cells with PBS.
2. Add 2–3 mL of pre-warmed 0.25% Trypsin-EDTA and incubate at 37°C until cells detach (2–10 minutes).
3. Neutralize trypsin by adding an equal volume of complete medium.
4. Centrifuge at 125xg for 5 minutes, aspirate the supernatant, and resuspend the cell pellet in fresh medium.
5. Seed cells at the appropriate density into new culture vessels and incubate under recommended conditions.

Cryopreservation Guidelines

• Cryopreservation Medium: Use Cryopreservation Medium (TM024) or complete growth medium supplemented with 10% DMSO.
• Freezing Protocol: Freeze at a controlled rate (-1°C per minute) before transferring to liquid nitrogen storage.

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Disclaimer

1. This product is intended for research use only and is not approved for diagnostic, therapeutic, or clinical applications.
2. Users are responsible for determining the suitability of this product for their specific application.
3. Cells must be handled under Biosafety Level 2 (BSL-2) containment following institutional guidelines.
4. No warranties are provided regarding performance beyond the described specifications.