Enzymatically prepared redox-responsive hydrogels as potent matrices for hepatocellular carcinoma cell spheroid formation.
Cellular spheroids have been received much attention in the biological and biomedical fields, especially as a base material for drug assays, regenerative medicine, and tissue engineering. Hydrogels have potential for scalable preparation of spheroids because they provide a spatial environment suitable for three-dimensional cell cultivation. Herein, the potential use of a redox-responsive hydrogel as a scaffold for preparation and recovery of spheroids is reported. A hydrogel composed of poly(ethylene glycol) (PEG), which can be degraded using cysteine as a reducing agent under mild conditions, is prepared by mixing an octa-thiolated PEG derivative (8-arm PEG-SH), horseradish peroxidase and a small phenolic compound (Glycyl-L-tyrosine). Human hepatocellular carcinoma cells (HepG2) are encapsulated in the hydrogel and cellular spheroids formed by proliferation within the scaffolds. After seven days of cultivation, the size of the HepG2 spheroids reached a diameter between ≈40 and 60 μm, depending on the 8-arm PEG-SH concentration. Liver-specific functions of the HepG2 spheroids such as albumin secretion and urea production are retained at higher levels than those of cells prepared from traditional two-dimensional mono layers. These results suggest that the system presented here has potential for preparation of cellular spheroids for tissue engineering applications.
Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Ibaraki, Japan.
Kousuke
K
Moriyama
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka, Japan.
Shono
S
Naito
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka, Japan.
Rie
R
Wakabayashi
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka, Japan.|Division of Biotechnology, Center for Future Chemistry, Kyushu University, Fukuoka, Japan.
Masahiro
M
Goto
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka, Japan.|Division of Biotechnology, Center for Future Chemistry, Kyushu University, Fukuoka, Japan.
Noriho
N
Kamiya
D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000628:therapy*; D064987:Cell- and Tissue-Based Therapy / Q000379:methods; D004151:Dipeptides / Q000737:chemistry; D056945:Hep G2 Cells; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D020100:Hydrogels / Q000737:chemistry / Q000627:therapeutic use; D008113:Liver Neoplasms / Q000473:pathology / Q000628:therapy*; D010084:Oxidation-Reduction; D011092:Polyethylene Glycols / Q000737:chemistry; D018874:Spheroids, Cellular / Q000737:chemistry* / Q000637:transplantation; D023822:Tissue Engineering
D004151:Dipeptides; D020100:Hydrogels; D011092:Polyethylene Glycols; C022013:glycyltyrosine; D006735:Horseradish Peroxidase
Hepatocytes; Horseradish peroxidase; Poly(ethylene glycol); Redox-responsive hydrogel; Spheroids