ArticleNature protocols2017
Synthesis and characterization of well-defined hydrogel matrices and their application to intestinal stem cell and organoid culture.
Article in Nature protocols, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 80 papers.
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Who cites it
80 citing papers in PubMed, 147 citations in OpenAlex.
- Organoids in Precision Radiotherapy: Methodological Foundations, Tumor-Specific Evidence, and Translational Roadmaps.Cancer medicine · 2026Review
- A Fully Defined GelMA-Based Matrix Allows Fine Tuning of Tissue-Relevant Biomechanical and Biochemical Cues for Organoid Culture.Advanced healthcare materials · 2026Article
- Advances in Biomaterial-Based Cardiac Organoids Engineering for Modeling Development and Congenital Heart Defects.ACS applied bio materials · 2026Review
- Synthetic PEG-4MAL Hydrogels Support Patient-Derived Human Intestinal Enteroid Culture.Cell biomaterials · 2026Article
- Stem cell-based tactics in the remodeling and treatment of intestinal diseases.Stem cell research & therapy · 2026Review
- Investigation of mechanical forces during multicellular early angiogenic sprouting by three-dimensional traction force microscopy in hydrogel matrices.Nature protocols · 2026Review
- From organoid culture to manufacturing: technologies for reproducible and scalable organoid production.npj biomedical innovations · 2026Review
- Controlling Intestinal Organoid Polarity using Synthetic Dynamic Hydrogels Decorated with Laminin-Derived IKVAV Peptides.Advanced healthcare materials · 2026Article
- 3D In Vitro Culture of Early Mouse Embryos and Trophoblast Tissue Explants.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Engineering Synthetic PEG Hydrogels to Model Peri-implantation Epiblast Morphogenesis with Tunable Biophysical Properties.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Invasin-functionalized PIC hydrogels enable long-term 3D culture of epithelial organoids.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Engineered basement membrane mimetic hydrogels to study mammary epithelial morphogenesis and invasion.Science advances · 2025Article
- The Reconstruction of Peripheral Auditory Circuit: Recent Advances and Future Challenges.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Review
- Mechanically and Chemically Defined PEG Hydrogels Improve Reproducibility in Human Cardioid Development.Advanced healthcare materials · 2025Article
- Mechanotransduction for therapeutic approaches: Cellular aging and rejuvenation.APL bioengineering · 2025Review
- Extracellular-Matrix-Mimetic Hydrogels by Using Nanomaterials.International journal of molecular sciences · 2025Review
- Review
- Review
- Engineering the 3D structure of organoids.Stem cell reports · 2025Review
- Article
20 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Growing cells within an extracellular matrix-like 3D gel is required for, or can improve, the growth of many cell types ex vivo. Here, we describe a protocol for the generation of well-defined matrices for the culture of intestinal stem cells (ISCs) and intestinal organoids. These matrices comprise a poly(ethylene glycol) (PEG) hydrogel backbone functionalized with minimal adhesion cues including RGD (Arg-Gly-Asp), which is sufficient for ISC expansion, and laminin-111, which is required for organoid formation. As such, the hydrogels present a defined and reproducible, but also tunable, environment, allowing researches to manipulate physical and chemical parameters, and examine their influence on ISC and organoid growth. Hydrogels are formed by an enzymatic cross-linking reaction of multiarm PEG precursors bearing glutamine- and lysine-containing peptides. PEG precursors containing either stable or hydrolytically degradable moieties are used to produce mechanically softening hydrogels, which are used for the expansion of ISCs or the formation of organoids, respectively. We also provide protocols for immunofluorescence analysis of cellular structures grown within these matrices, as well as for their dissociation and retrieval of cells for downstream use. Hydrogel precursors can be produced and their mechanical properties characterized to ascertain stiffness within 5-7 d. Hydrogel formation for ISC expansion or organoid formation takes 1-2 h. The materials described here can be readily adapted for the culture of other types of normal or transformed organoid structures.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.