HepG2 Cell Line Origin and Characteristics
HepG2 is an immortalized cell line consisting of human liver carcinoma cells, derived from the liver tissue of a 15-year-old Caucasian male who had a well-differentiated hepatocellular carcinoma, which is the fifth most-common cancer worldwide. The HepG2 cell line is commonly used in drug metabolism and hepatoxicity studies. HepG2 cells exhibit an epithelial-like morphology with a modal chormosome number of 55. They are also non-tumorigenic and have high proliferation rates.
HepG2 cells have adherent properties and grow as monolayers in small aggregates. HepG2 can be grown successfully at a large scale and stimulated with human growth hormone. They are also capable of secreting many plasma proteins, such as transferrin, fibrinogen, plasminogen and albumin. Additionally, HepG2 cells are an ideal in vitro model to study the intracellular dynamics of cell surface domains, such as bile canaliculi and sinusoidal membrane proteins and lipids in human hepatocytes. Studying such dynamics contribute to better understanding certain liver diseases that result from an incorrect cell surface protein distribution.
HepG2 Cell Culturing Protocol
HepG2 complete medium
Eagle’s Minimum Essential Medium (EMEM) supplemented with 10% FBS; DMEM and RPMI1640 are also alternatives that work well. Aspirate and add fresh culture medium every 2-3 days. HepG2 cell doubling time is 48 hours.
- To passage cells, rinse cell monolayer with 1x PBS twice and add pre-warmed (37°C) 0.05% Trypsin-EDTA solution to cover the bottom of the flask; incubate for 5 – 7 minutes
- As cells detach, neutralize the Trypsin by adding 4x volume of complete growth medium with 10% FBS and gently resuspend the cells by pipetting
- To avoid clumping do not agitate the cells by shaking the flask while waiting for detachment
- Split cells 1:4 every 3 days or 1:8 every 6 days
- Cultures should be incubated at 37°C in a humidified atmosphere with 5% CO2
Subculture Troubleshooting Procedures
Low cell viability after passaging:
- Dissociation agent left on cells too long; only expose cells to dissociation agent long enough for cell detachment
- Pipette gently during passaging procedures; cells are fragile when exposed to dissociation agents
Cells are difficult to detach:
- Cell-to-cell junctions are tight due to cell growth being 100% confluent and dissociation agent cannot reach cell interface; subculture cells before confluent
- Use higher concentration of dissociation agent; incubate flask at 37°C to increase enzymatic activity
- Wash flask twice with sterile 1x PBS prior to addition of the dissociation agent
Clumps form after detachment:
- Cells were centrifuged too fast; do not spin cells faster than 100 x g to pellet
- Place the flask or vial on ice to decrease aggregation before use
HepG2 Cell Line Derived Xenograft
HepG2 cells are inoculated in immunocompromised mice to create the HepG2 Cell Line Derived Xenograft (CDX) mouse model. The HepG2 xenograft of human hepatocellular carcinoma (HCC) enables studies targeting antiangiogenesis (i.e. rapamycin, bevacizumab) or tumor growth inhibition (e.g. sorafenib).
Stable Cell Line Generation
HepG2 cells have been demonstrated to be Neomycin G418 resistant (400 µg/mL). Development of HepG2 stable cell line services are provided by Altogen Labs CRO
*NOTE: All IP rights to the HepG2 cell line belongs solely to the Wistar Institute.
HepG2 Resources
siRNA Delivery – In Vivo Transfection Kits
CRO Pre-clinical Research Services: Xenograft animal models
Generation of Stably Expressing Cell Lines in 28 Days
Encapsulation of Protein, RNA, mRNA, and DNA Molecules into Liposomes
HepG2 cells Forum: Research methods & Laboratory techniques: Link
HepG2 Cell Culture | HepG2 Cell Transfection | HepG2 Sable Cell Line | Get HepG2 Transfection Reagent