Glycogen storage assay

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Scope of the method

The Method relates to
  • Human health
The Method is situated in
  • Basic Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Animal derived cells / tissues / organs
Species from which cells/tissues/organs are derived
rat, mice, humans
Type of cells/tissues/organs
parenchymal liver cells, stem cell-derived hepatocyte-like cells

Description

Method keywords
  • glycogen
  • liver
Scientific area keywords
  • liver research
  • liver disease
  • inborn error of metabolism
Method description

Cultivated liver cells are fixed with 4% (w/v) paraformaldehyde (PFA) for 10 minutes at room temperature and subsequently incubated for 15 minutes with 100 mM glycin solution, used to saturate reactive groups generated after PFA fixation. These cells are subsequently incubated for 10 minutes with 1% (w/v) Periodic Acid Reagent and 15 minutes of Schiff's Reagent to stain glycogen. Finally, the nuclei are counterstained with haematoxylin solution.

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Lab equipment

Fume hood.

Method status
  • History of use
  • Internally validated
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages

Easy assay to investigate the capacity of hepatocyte-like cells to store glycogen.

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

Contact person

Joery De Kock

Organisations

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