HepG2 Culture/Medium    HepG2 Stable Line

HepG2 Cells

human liver hepatocellular carcinoma cell line

Cell culture protocols, stable and transient transfection

HepG2 are adherent, epithelial-like cells growing as monolayers and in small aggregates, have a model chromosome number of 55. HepG2 cell line was derived from the liver tissue of fifteen year old male with differentiated hepatocellular carcinoma. Not tumorigenic in immunocompromised mice. Secrete plasma proteins, such as albumin, transferrin, fibrinogen, a-2-macroglobulin, plasminogen. Cells respond to stimulation with HGH.

Cytogenetics: human hyperdiploid karyotype - 52(47-54)<2n>XY, +2, +14, +17, +20, +2mar, t(1;21) (p22.2;p11-12), i(17q)/der(17)t(17;17)(p11;q11)


Copyright picture from altogen.com. Reproduced with permission from Altogen Biosystems.

Culturing protocol:

1. Use Eagle's Minimum Essential Medium (DMEM and RPMI1640 also works well) supplemented with 10% FBS. Discard culture medium every 2-3 days
2. To passage cells, briefly rinse cell monolayer with 1xPBS twice and add pre-warmed (37°C) 0.05% Trypsin-EDTA solution for 5 - 7 minutes
3. Once cell layer is dispersed (5-7 min at 37°C) deactivate Trypsin by adding equal volume of complete growth medium with 10% FBS
4. To avoid clumping don't agitate the cells by shaking the flask while waiting for the cells to detach
5. Split cells 1:4 every 3 days (recommended), or 1:8 every 6 days.
6. Cultures should be incubated at 37°C in humidified atmoshere with 5% CO2

Stable Cell Line Generation:

HepG2 cells have been demonstrated to be Neomycin G418 resistant (400 µg/mL).


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