Generation of Hepatic Stellate Cells from Human Pluripotent Stem for in vitro liver fibrosis studies

Scope of the method

The Method relates to
  • Human health
The Method is situated in
  • Basic Research
  • Translational - Applied Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Human derived cells / tissues / organs
Specify the type of cells/tissues/organs
Non-tumor liver tissue, Non-tumor cirrhotic liver tissue

Description

Method keywords
  • Pluripotent stem cells
  • Hepatic stellate cells
  • organoids
  • Liver spheroids
  • In vitro liver model
  • Non-parenchymal cells
  • HepaRG
Scientific area keywords
  • Liver fibrosis
  • Disease modelling
  • Toxicity assessment
  • hepatocytes
Method description

We established a protocol to efficiently generate hepatic stellate cells (HSCs) from human pluripotent stem cells (PSCs). Our procedure generated complex in vitro spheroid cultures that better mimic the complexity of the liver as well as liver function. In co-culture, iPSC-HSCs promote maintenance of hepatocyte metabolic functionality while being able to respond to hepatocyte-mediated toxicity, activating and promoting intra-spheroid fibrogenesis, one of the main drug-associated adverse liver outcomes. iPSC-HSCs display functional and phenotypic features of human primary cultured HSCs, indicating that they may be a highly suitable cell source of human HSCs for culture-based studies.

Lab equipment
  • - Incubator,
  • - Cell culture hood, 
  • - Flow cytometer,
  • - Laser Scanning Confocal microscope.
Method status
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages
  • - Protocol is highly robust,
  • - Yields 70%–80% iPSC-HSCs,
  • - Highly reproducible.
Challenges

In 2D the responsive of iPSC-HSCs to external signals is rather limited. Thus far, the method has been used successfully in 3 different institutes using 3 different hESC/hIPSC cell lines, but more should be tested.

Modifications

Higher throughput and better quality control for the different stages of hiPSC to HSC differentiations.

Future & Other applications

Can be used for several applications, such as developmental studies, fibrosis modeling, drug screening, liver spheroid generation, and, eventually, regenerative medicine.

References, associated documents and other information

References

Coll, Mar et al. Generation of Hepatic Stellate Cells from Human Pluripotent Stem Cells Enables In Vitro Modeling of Liver Fibrosis. Cell Stem Cell, Volume 23, Issue 1, 101 - 113.e7

Associated documents
Links
Liver cell biology research group

Contact person

Leo A. van Grunsven

Organisations

Vrije Universiteit Brussel
Liver Cell Biology research group
Belgium
Brussels Region

Partners

KU Leuven