Fabrication of Inverted Colloidal Crystal Poly(ethylene glycol) Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering
Open accessAbstract
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.
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Cell-binding cues from collagen, fibronectin, or laminin have frequently been added to synthetic hydrogels to allow for organoid growth and differentiation.
“Cell-binding cues from collagen, fibronectin, or laminin have frequently been added to synthetic hydrogels to allow for organoid growth and differentiation.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [159]
Attachment of human iPSCs was successful in assemblies functionalized with collagen I and laminin-521 but not with fibronectin.
“Attachment of human iPSCs was successful in assemblies functionalized with collagen I and laminin-521 but not with fibronectin.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [159]
Ng and colleagues created functional human liver organoids derived from human iPSCs in a colloidal crystal of PEG functionalized with collagen I, fibronectin, or laminin-521.
“Ng and colleagues created functional human liver organoids derived from human iPSCs in a colloidal crystal of PEG functionalized with collagen I, fibronectin, or laminin-521158,159.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [159]
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Towards organoid culture without Matrigel
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- Authors
- Shirahama, H., Kumar, S. K., Jeon, W.-Y., Kim, M. H., Lee, J. H., Ng, S. S., Tabaei, S. R., & Cho, N.-J.
- Affiliations
- Nanyang Technological University (SG)
- ORCID
- Venue
- Journal of Visualized Experiments
- Publisher
- MyJove Corporation
- Published
- 2016-08-27
- Issue
- 114
- ISSN
- 1940-087X (ISSN-L)
- PMC
- PMC5091954
- OpenAlex
- W2517160664
- Cited by
- 14
- References
- 62
- Citations by year
- 2024: 1 · 2022: 2 · 2021: 2 · 2020: 2 · 2019: 3 · 2018: 3
- FWCI
- 0.96
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Visualized Experiments (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Research Foundation (NRF-CRP10-2012-07, NRFF2011-01)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Hydrogels: synthesis, properties, applications
- Keywords
- poly(ethylene glycol) diacrylate, particulate leaching, 3D cell culture, liver tissue engineering, hepatocyte function, Huh7.5 cells, extracellular matrix coating
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