Fabrication of Inverted Colloidal Crystal Poly(ethylene glycol) Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering.
The ability to maintain hepatocyte function in vitro, for the purpose of testing xenobiotics' cytotoxicity, studying virus infection and developing drugs targeted at the liver, requires a platform in which cells receive proper biochemical and mechanical cues. Recent liver tissue engineering systems have employed three-dimensional (3D) scaffolds composed of synthetic or natural hydrogels, given their high water retention and their ability to provide the mechanical stimuli needed by the cells. There has been growing interest in the inverted colloidal crystal (ICC) scaffold, a recent development, which allows high spatial organization, homotypic and heterotypic cell interaction, as well as cell-extracellular matrix (ECM) interaction. Herein, we describe a protocol to fabricate the ICC scaffold using poly (ethylene glycol) diacrylate (PEGDA) and the particle leaching method. Briefly, a lattice is made from microsphere particles, after which a pre-polymer solution is added, properly polymerized, and the particles are then removed, or leached, using an organic solvent (e.g., tetrahydrofuran). The dissolution of the lattice results in a highly porous scaffold with controlled pore sizes and interconnectivities that allow media to reach cells more easily. This unique structure allows high surface area for the cells to adhere to as well as easy communication between pores, and the ability to coat the PEGDA ICC scaffold with proteins also shows a marked effect on cell performance. We analyze the morphology of the scaffold as well as the hepatocarcinoma cell (Huh-7.5) behavior in terms of viability and function to explore the effect of ICC structure and ECM coatings. Overall, this paper provides a detailed protocol of an emerging scaffold that has wide applications in tissue engineering, especially liver tissue engineering.
School of Materials Science and Engineering, Nanyang Technological University.
Hitomi
H
Shirahama
School of Materials Science and Engineering, Nanyang Technological University.
Supriya K
SK
Kumar
School of Materials Science and Engineering, Nanyang Technological University.
Won-Yong
WY
Jeon
School of Materials Science and Engineering, Nanyang Technological University.
Myung Hee
MH
Kim
School of Materials Science and Engineering, Nanyang Technological University.
Jae Ho
JH
Lee
School of Materials Science and Engineering, Nanyang Technological University.
Soon Seng
SS
Ng
School of Materials Science and Engineering, Nanyang Technological University.
Seyed R
SR
Tabaei
School of Materials Science and Engineering, Nanyang Technological University; School of Chemical and Biomedical Engineering, Nanyang Technological University; njcho@ntu.edu.sg.
Nam-Joon
NJ
Cho
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