Isolation and cultivation of rat liver epithelial cells

Commonly used acronym: rLEC

Scope of the method

Alternative method relates to
  • Human health
Alternative method is situated in
  • Basic Research
Type of alternative method
  • In vitro - Ex vivo
This method makes use of
  • Animal derived cells / tissues / organs
Species from which cells/tissues/organs are derived
Rattus norvegicus
Type of cells/tissues/organs
rat liver epithelial cells

Description

Method keywords
  • liver
  • epithelial cells
  • isolation
Scientific area keywords
  • liver research
  • liver cells
Method description

Rat liver epithelial cells (rLEC) can be isolated from 8-day old male Sprague-Dawley rats. Briefly, small fragments of neonatal rat livers are incubated for 15 minutes with 4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid (HEPES) buffered trypsin solution [0.25% (v/v)] and washed twice with calcium- and magnesium-free phosphate-buffered saline (PBS) before plating. Elimination of contaminating fibroblasts is accomplished by taking advantage of their faster attachment to plastic culture dishes (plate-and-wait method). Growth medium consisted of Williams’ E medium without glutamine, 10 % (v/v) fetal bovine serum (FBS), 0.68 mM L-glutamine, 50 µg/mL streptomycin sulphate, 7.33 IU/mL benzyl penicillin, 50 µg/mL kanamycin monosulphate and 10 µg/mL sodium ampicillin. Cell cultures are incubated at 37 °C in a 5 % CO2 and 95 % air humidified atmosphere. Growth media is changed completely every 2 days.

Lab equipment
  • Biosafety cabinet level 1 ;
  • Cell incubator ;
  • Centrifuge.
Method status
  • History of use
  • Internally validated
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages

Robust isolation and cultivation method for rat liver epithelial cells.

References, associated documents and other information

References

De Kock J, Snykers S, Branson S, Jagtap S, Gaspar JA, Sachinidis A, Vanhaecke T, Rogiers V. (2012) A liver-derived rat epithelial cell line from biliary origin acquires hepatic functions upon sequential exposure to hepatogenic growth factors and cytokines. Curr Med Chem. 19(26):4523-33

Associated documents

Contact person

Joery De Kock

Organisations

Vrije Universiteit Brussel
Pharmaceutical and Pharmacological Sciences (FARM)
In Vitro Toxicology and Dermato-cosmetology
Belgium
Brussels Region