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Galactosylated reversible hydrogels as scaffold for HepG2 spheroid generation
Yuhan Wu1, Ziqi Zhao1, Ying Guan1
1State Key Laboratory of Medicinal Chemical Biology and Key Laboratory of Functional Polymer Materials, Institute of Polymer Chemistry, College of Chemistry, Nankai University, and Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300071, China.
Researchers developed a novel, reversible galactosylated hydrogel scaffold for hepatocyte culture. This scaffold supports cell viability, promotes spheroid formation, and allows for easy spheroid harvesting, advancing liver tissue engineering.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Hepatocyte Culture
Background:
- Galactosylated scaffolds enhance hepatocyte viability, spheroid formation, and liver-specific functions.
- Harvesting spheroids from 3D scaffolds for analysis remains challenging.
Purpose of the Study:
- To develop a novel, reversible galactosylated hydrogel scaffold for improved hepatocyte culture and spheroid retrieval.
- To investigate the effect of galactose ligand content on cell viability, spheroid formation, and liver-specific functions.
Main Methods:
- Fabrication of a thermosensitive hydrogel scaffold using poly(N-isopropylacrylamide) (PNIPAM) microgel and poly(ethylene glycol) (PEG).
- Incorporation of galactosylated PNIPAM microgel to create scaffolds with varying galactose ligand densities.
- Encapsulation of HepG2 cells within the in situ-formed hydrogels and assessment of cell viability, spheroid formation, albumin secretion, and urea synthesis.
Main Results:
- The hydrogel scaffold solidifies at physiological temperature and liquefies upon cooling, enabling easy spheroid harvesting.
- Increased galactose ligand content correlated with enhanced HepG2 cell viability.
- Galactose-containing scaffolds promoted the formation of compact multicellular spheroids and maintained higher levels of liver-specific functions (albumin secretion, urea synthesis).
Conclusions:
- The reversible galactosylated hydrogel scaffold successfully supports hepatocyte culture, promotes spheroid formation, and facilitates spheroid retrieval.
- This novel scaffold offers a promising platform for large-scale fabrication and application of hepatocyte spheroids in liver research and regenerative medicine.
