HepG2 cell culture, stable transfection protocol

HepG2 is a perpetual 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. Hepatocellular carcinoma is, worldwide, the fifth most-common cancer. The morphology of HepG2 cells is epithelial and they have 55 chromosome pairs. HepG2 cells can be grown successfully at a large scale, and secret many plasma proteins, such as transferring, fibrinogen, plasminogen and albumin. They can be stimulated with human growth hormone. HepG2 are adherent, epithelial-like cells growing as monolayers and in small aggregates. Not tumorigenic in immunocompromised mice.

HepG2 transfection kit is commercially available from Altogen Biosystems

hepg2w

hepg2cells

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

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)

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). Development of HepG2 stable cell line services provided by Altogen Labs CRO

HepG2 Resources:

HepG2 Transfection Kit

Xenograft Models

siRNA Delivery – In Vivo Transfection Kits

CRO Pre-clinical Research Services: Xenograft animal models

HepG2 Cell Banking services

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