Human liver spheroid co-cultures to investigate parenteral nutrition-induced hepatotoxicity

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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
C3A cells (clonal derivative of the human hepatoma HepG2 cell line), LX-2 cells (immortalized activated human hepatic stellate cell line)

Description

Method keywords
  • Liver spheroids
  • co-culture
  • RNA sequencing
  • Hepatotoxicity
  • spheroids
  • In vitro liver model
  • 3D in vitro model
  • liver cells
Scientific area keywords
  • Total Parenteral Nutrition
  • Instestinal failure-associated liver disease
  • liver injury
  • in vitro toxicology
  • TPN
  • IFALD
  • nutrition
Method description

Total Parenteral Nutrition (TPN) can cause adverse effects, including metabolic disorders and liver injury. TPN-associated liver injury, known as intestinal failure-associated liver disease (IFALD), represents a significant problem affecting up to 90% of individuals receiving TPN. Despite numerous animal studies and clinical observations, the molecular mechanisms driving IFALD remain largely unknown. For this, a three-dimensional (3D) spheroid co-culture system consisting of both human parenchymal and non-parenchymal liver cells was used to elucidate the mechanisms of TPN-associated liver injury. Human liver spheroid co-cultures were set up using C3A cells, a clonal derivative of the human hepatoma HepG2 cell line, and LX-2 cells, an immortalized activated human hepatic stellate cell line.

Lab equipment
  • - Laminar air flow,
  • - Microscope, 
  • - SpectraMax iD3 Multi-Mode Reader,
  • - Spectrophotometer, 
  • - Attune Acoustic Focusing Cytometer.
Method status
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages

The use of human-centered in vitro systems can overcome the limitations of clinical research and animal experimentation in the IFALD field, and allows in-depth investigation at the mechanistic level.

Challenges

Despite providing important molecular and cellular insights into mechanisms of liver injury secondary to TPN exposure, a limitation of the present study is the lack of direct clinical translation. This is particularly reflected in attempts to correlate the dosages of TPN used in vitro to those observed in clinics during a TPN regimen.

Future & Other applications

Computational toxicology methods that predict chemical toxicity and correlate in vitro and in vivo concentrations are available and could aid in the efforts needed to increase clinical relevance. In particular, physiologically based pharmacokinetic (PBPK) modeling can facilitate quantitative in vitro to in vivo extrapolation (QIVIVE) and allow to combine in silico and in vitro parameters and convert in vitro concentration–response curves into relevant in vivo exposures.

A combination of advanced heterotypic cell models, suitable in vitro and omics analysis, systems toxicology approaches, including AOPs and their networks and PBPK modeling holds great promise for advancing the translational research oriented toward unraveling further IFALD mechanisms and encouraging the safe use of TPN.

References, associated documents and other information

Links
Investigation of parenteral nutrition-induced hepatotoxicity using human liver …
IVTD Research team (VUB)

Organisations

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