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Related Experiment Video

Updated: Jun 22, 2025

Decellularization and Recellularization of Whole Livers
09:24

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Published on: February 4, 2011

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Liver click dECM hydrogels for engineering hepatic microenvironments.

Laura A Milton1, Jordan W Davern2, Luke Hipwood3

  • 1Faculty of Engineering, School of Mechanical, Medical and Process Engineering, Queensland University of Technology, Brisbane, Australia; Centre for Biomedical Technologies, Queensland University of Technology, Brisbane, Australia; Gelomics Pty Ltd, Brisbane, Australia.

Acta Biomaterialia
|July 3, 2024
PubMed
Summary
This summary is machine-generated.

We developed new click hydrogels from liver extracellular matrix (dECM) for better 3D cell cultures. These reproducible dECM hydrogels offer precise mechanical control and enhanced liver cell function for drug testing.

Keywords:
3D cell cultureDecellularized extracellular matrixHydrogelLiverMichael-type addition

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Cell Biology

Background:

  • Decellularized extracellular matrix (dECM) hydrogels offer native tissue microenvironments for 3D cell cultures but suffer from poor reproducibility due to uncontrolled collagen crosslinking.
  • Existing dECM hydrogels lack precise control over mechanical properties, limiting their utility for modeling diverse physiological and pathological conditions.
  • Developing reproducible and tunable dECM-based biomaterials is crucial for advancing in vitro cell culture models.

Purpose of the Study:

  • To create a reproducible and mechanically tunable liver dECM hydrogel system using click chemistry.
  • To optimize decellularization protocols for maximal ECM retention and DNA removal.
  • To evaluate the cytocompatibility and functional performance of the novel dECM hydrogels in supporting liver cells.

Main Methods:

  • Optimized liver tissue decellularization using Triton X-100 and ammonium hydroxide.
  • Functionalized pepsin-solubilized liver dECM with thiols (dECM-SH) via EDC/NHS coupling with L-Cysteine.
  • Formed hydrogels through Michael-type addition reaction between dECM-SH and 4-arm PEG-maleimide.
  • Characterized hydrogel mechanical properties (Young's moduli 1-7 kPa) and assessed liver cell (HepG2, HepaRG) viability, proliferation, and function in 3D culture.

Main Results:

  • Achieved near-complete DNA removal and preserved native liver proteome during decellularization.
  • Synthesized optically clear, covalently crosslinked liver dECM hydrogels with tunable mechanical properties.
  • Demonstrated excellent cytocompatibility, supported HepG2 and HepaRG cell growth, and significantly enhanced liver-specific functions (metabolic activity, CYP enzyme activity, excretion) compared to controls.
  • Click dECM-SH hydrogels exhibited superior handling and reproducibility compared to traditional dECM hydrogels.

Conclusions:

  • Thiolated liver dECM hydrogels formed via click chemistry provide a highly controlled and reproducible platform for 3D liver cell culture.
  • These novel hydrogels closely mimic native liver ECM properties and enable precise tuning of mechanical characteristics.
  • The enhanced liver-specific cellular functions observed in these hydrogels represent a significant advancement for in vitro liver models in drug discovery and toxicology.