Drug-induced phospholipidosis in hepatic cells derived from human skin-derived precursors

Scope of the method

The Method relates to
  • Human health
The Method is situated in
  • Translational - Applied Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Human derived cells / tissues / organs

Description

Method keywords
  • toxicology
  • in vitro
  • drug testing
  • Intracellular lipids
  • phospholipids
  • Phospholipidosis
  • amiodarone
Scientific area keywords
  • hepatic differentiation
  • Hepatotoxicity
  • skin-derived precursors
  • skin stem cells
Method description

Drug-induced phospholipidosis (DIPL) is a metabolic disorder characterized by an excessive intracellular accumulation of phospholipids caused by cationic drugs. Hepatic cells derived from human skin are evaluated as an in vitro model to investigate DIPL and its mechanisms. Human skin stem cells (hSKP) are isolated, under informed consent, from human circumcised foreskin samples of young boys and hSKP are differentiated for 24 days to obtain hepatic-like cells (hSKP-HPC), as previously described. hSKP-HPC are exposed to amiodarone, a drug known to induce phospholipidosis in humans. Upon exposure to amiodarone for 24, 48, 72h, hSKP-HPC retain intracellular phospholipids, form lamellar bodies and show alterations at the gene expression level. Overall, these findings prove that hSKP-HPC might contribute to setting up an accurate in vitro platform for hepatotoxicity testing.

Lab equipment
  • Laminar air flow ;
  • Flow cytometry ;
  • Transmission electron microscopy ;
  • Reverse transcriptase-polymerase chain reaction (qPCR) reagents.
Method status
  • History of use

Pros, cons & Future potential

Advantages
  • Applicability of hSKP-HPC for the quick assessment of drug-induced phospholipidosis in vitro ;
  • Different human donors can be tested to assess toxicity.
Future & Other applications

In vitro toxicity testing in the drug development process

References, associated documents and other information

References

W. H. Halliwell, “Amphiphilic Drug-Induced Phospholipidosis,” Toxicol. Pathol., vol. 25, no. 1, pp. 53–60, 1997. [2] R. M. Rodrigues et al., “Human skin-derived stem cells as a novel cell source for in vitro hepatotoxicity screening of pharmaceuticals.,” Stem Cells Dev., vol. 23, no. 1, pp. 44–55, 2014

Associated documents
Article A Natale 2017.pdf

Organisations

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