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Squaramide-Based Supramolecular Materials Drive HepG2 Spheroid Differentiation.

Tingxian Liu1, Linda van den Berk2, Joeri A J Wondergem3,4

  • 1Department of Supramolecular and Biomaterials Chemistry, Leiden Institute of Chemistry, Leiden University, P.O. Box 9502, Leiden, 2300 RA, Netherlands.

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Summary
This summary is machine-generated.

A new synthetic squaramide hydrogel matrix effectively matures HepG2 liver cells in 3D culture. This advanced material enhances drug toxicity screening by improving cell function compared to traditional Matrigel.

Keywords:
3D cell cultureHepG2 spheroidshydrogel​ssquaramidesupramolecular

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Area of Science:

  • Biomaterials Science
  • Hepatocyte Cell Culture
  • Drug Discovery and Toxicology

Background:

  • HepG2 cell 2D cultures lack maturity for drug toxicity screening.
  • While 3D Matrigel cultures improve some liver markers, they still lack primary hepatocyte functions.
  • Controlled synthetic matrices are needed to enhance HepG2 spheroid maturation.

Purpose of the Study:

  • To develop a fully synthetic, minimalistic supramolecular matrix for improved HepG2 spheroid culture.
  • To investigate the potential of squaramide-based hydrogels to promote HepG2 cell differentiation and function.
  • To compare the efficacy of the synthetic matrix against Matrigel for drug toxicity screening applications.

Main Methods:

  • A modular approach using squaramide synthons and RGD peptides to create supramolecular hydrogels.
  • Co-assembly of functionalized and native monomers to form soft, self-recovering hydrogels with tunable RGD concentration.
  • Culture of HepG2 spheroids within the hydrogels for 21 days and assessment of cell viability, differentiation, and gene/protein expression.

Main Results:

  • Formation of stable HepG2 spheroids (≈150 µm) with high cell viability in the squaramide hydrogels.
  • Significantly enhanced mRNA and protein expression of phase I/II metabolic enzymes, drug transporters, and liver markers compared to Matrigel.
  • Demonstrated synthetic accessibility and effectiveness of the squaramide hydrogels for HepG2 differentiation.

Conclusions:

  • Fully synthetic squaramide hydrogels are effective for culturing and maturing HepG2 spheroids.
  • This supramolecular matrix significantly improves HepG2 cell differentiation and function over Matrigel.
  • Squaramide hydrogels show potential to replace natural matrices in high-throughput drug toxicity screening.