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Collagen Vitrigel: A Novel Scaffold that can Facilitate a Three-Dimensional Culture for Reconstructing Organoids
Open accessAbstract
Three-dimensional reconstructed organoids in vitro are valuable for not only regenerative medicine but also drug development. However, the manipulation of conventional three-dimensional cultures is not simple. We describe a nylon membrane ring-embedded or a pressed silk sheet-embedded scaffold made of collagen “vitrigel” that can facilitate three-dimensional cultures for reconstructing an epithelial-mesenchymal model or a hard connective tissue model, respectively. Here we define vitrigel as a gel in a stable state produced by rehydration after the vitrification of a traditional hydrogel. The collagen vitrigel was successfully prepared in three steps involving a gelation process in which a cold and clear neutral salt solution containing type I collagen formed an opaque and soft gel by incubation at 37°C, a vitrification process in which the gel becomes a rigid material like glass after sufficient drying out, and finally a rehydration process to convert the vitrified material into a thin and transparent gel membrane with enhanced gel strength. The framework-embedded collagen vitrigel scaffold that can be easily reversed by forceps was prepared by inserting a nylon ring or a silk sheet in the collagen solution prior to the gelation. The scaffold enabled culturing anchorage-dependent cells on both surfaces of the collagen vitrigel by the manipulation of two-dimensional cultures and consequently resulted in reconstructing a three-dimensional organoid. An intestinal epithelial-mesenchymal model was reconstructed by coculturing fibroblasts on the opposite side of monolayered Caco-2 cells on the nylon ring-embedded collagen vitrigel. Also, fibroblasts seeded on both surfaces of the silk sheet-embedded collagen vitrigel proliferated well and formed multilayers and some cells invaded into the vitrigel framed by the network of numerous strong silk filaments, suggesting a reconstruction of a hard connective tissue model. These data demonstrate that the collagen vitrigel is a valuable scaffold for tissue engineering.
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“Other examples are vitrified collagen I for human intestinal organoids86 and murine renal organoids87, as well as fibrin supplemented with laminin-111 capable of supporting various murine and human epithelial organoids88.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [86]
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Metadata
- Authors
- Takezawa, T., Ozaki, K., Nitani, A., Takabayashi, C., & Shimo-Oka, T.
- Affiliations
- Institute of Agrobiological Sciences (JP)
- Laboratory of Animal Cell Biology, National Institute of Agrobiological Sciences, Ikenodai 2, Tsukuba, Ibaraki 305-0901, Japan
- Corresponding author
- Takezawa, Toshiaki
- ORCID
- Takezawa, Toshiaki
- Venue
- Cell Transplantation
- Publisher
- SAGE Publications
- Published
- 2004-05
- Volume
- 13
- Issue
- 4
- Pages
- 463-474
- ISSN
- 0963-6897 (ISSN-L)
- 1555-3892
- OpenAlex
- W2031151230
- Cited by
- 165
- References
- 36
- Citations by year
- 2026: 5 · 2025: 4 · 2024: 5 · 2023: 11 · 2022: 10 · 2021: 14 · 2020: 6 · 2019: 7 · 2018: 11 · 2017: 6 · 2016: 13 · 2015: 9 · 2014: 10 · 2013: 12 · 2012: 5
- FWCI
- 1.54
- Type
- journal article
- Language
- en
- Open access
- green
- doaj.org
- Locations
- Cell Transplantation (publishedVersion)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Topics
- Physical Sciences › Materials Science › Biomaterials › Hydrogels: synthesis, properties, applications
- Physical Sciences › Materials Science › Biomaterials › Collagen: Extraction and Characterization
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- collagen scaffolds, vitrification, Caco-2 cells, fibroblast invasion, tissue engineering scaffolds
- MeSH terms
- Cells, Cultured, Collagen, Connective Tissue, Epithelium, Fibroblasts, Gels, Humans, Mesoderm, Coculture Techniques, Cell Culture Techniques, Caco-2 Cells, Tissue Engineering
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