Product Details

• Catalog Number: T0063
• Unit Size: 1×10^6 cells / 1.0 ml
• Species: Human (Homo sapiens)
• Tissue of Origin: Liver
• Cell Type: Immortalized Hepatocytes
• Growth Properties: Adherent, epithelial-like
• Immortalization Method: Lentiviral transduction with HPV E6/E7, hTERT & MycT58A
• Expression Markers: Beta-2 microglobulin
• Biosafety Level: BSL-2
• Storage Conditions: Below -130°C (liquid nitrogen vapor phase)
• Shipping Conditions: Shipped on dry ice
• Format: Cryopreserved frozen cells

Overview

The Immortalized Human Hepatocytes (IHH) offer a superior alternative to primary hepatocytes, ensuring consistent performance in liver-related studies. Engineered for extended proliferation while maintaining key hepatic functions, they provide an excellent platform for research in drug metabolism, toxicity assessment, and liver cell biology.

Key Features and Benefits

• • • Stable Hepatic Model: Provides a consistent and reproducible system for liver-based research.
    • Enhanced Proliferation: Overcomes limitations of primary hepatocytes with long-term culture capabilities.
    • Drug Screening & Toxicology: Ideal for pharmaceutical and toxicological investigations.
    • Supports 3D Culture & Co-Culture: Enables advanced hepatocyte applications in tissue engineering and disease modeling.
    • Expression of Hepatic Markers: Retains key metabolic and transport properties of liver cells.

Culture & Handling Guidelines

Recommended Culture Conditions

• Coating: PriCoat™ ECM T25 Flasks (G999) or Applied Cell Extracellular Matrix (G422) for optimal adhesion.
• Growth Medium: PriGrow IX (TM019) supplemented with:
  • 10% Fetal Bovine Serum (FBS)
  • 1% Penicillin/Streptomycin Solution (G255)
• Incubation Conditions: 37°C, 5% CO₂ humidified atmosphere
• Seeding Density: 30,000 cells/cm²
• Doubling Time: 30–35 hours

Thawing Protocol

1. Quickly thaw the vial in a 37°C water bath for no more than 2 minutes. Ensure the vial cap remains above water level to minimize contamination.
2. Decontaminate the vial by spraying with 70% ethanol and transfer it to a biological safety cabinet.
3. Transfer the cell suspension into a sterile 15mL conical tube containing 5mL of pre-warmed complete growth medium.
4. Centrifuge at 125xg for 5–7 minutes.
5. Aspirate the supernatant and resuspend the cell pellet in fresh complete medium. Seed the cells into a pre-coated T25 flask and incubate under recommended conditions.

Subculturing Guidelines

1. Remove 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 (typically 2–10 minutes).
3. Neutralize the trypsin with an equal volume of complete medium.
4. Centrifuge at 125xg for 5 minutes. Aspirate the supernatant and resuspend the pellet in fresh medium.
5. Seed the cells at the desired density into new culture vessels and incubate under recommended conditions.

Cryopreservation Guidelines

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

Related Products

• Recombinant Human TGF Beta-1 (TGFB1) – Z101555
• Recombinant Human IGF1 (E. coli) – Z100385
• Recombinant Human EGF – Z100139

Disclaimer

• This product is intended solely for research purposes and is not approved for human or animal therapeutic use.
• All test parameters are based on standard laboratory conditions. Performance may vary depending on user protocols.
• Users are responsible for ensuring compliance with local regulations regarding the handling and disposal of biological materials.

References

1. Blackman, Marine CNM, et al. “Mitochondrial protein Cox7b is a metabolic sensor driving brain-specific metastasis of human breast cancer cells.” Cancers 14.18 (2022): 4371.