ArticleBio-protocol2022
From 3D to 2D: Harmonization of Protocols for Two-dimensional Cultures on Cell Culture Inserts of Intestinal Organoids from Various Species.
Article in Bio-protocol, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
What it found
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Who cites it
9 citing papers in PubMed, 13 citations in OpenAlex.
- Nanofiber-Supported Decellularized Matrix Membrane Creates a Biomimetic Biochemical and Biophysical Niche for Intestinal Organoid Epithelium.Advanced healthcare materials · 2026Article
- Long-term feeder cell-free cat intestinal organoid cultures to study Toxoplasma gondii's sexual development.Communications biology · 2026Article
- Infectious Diarrhea in Early Childhood across the Global South: Etiologic Diversity and Pathogenic Mechanisms.ACS infectious diseases · 2026Review
- Establishing and Maintaining 3D Organoid Cultures From Human Fallopian Tube Epithelium.Bio-protocol · 2025Article
- Article
- 3DFrontiers in pharmacology · 2025Review
- Comparison of Glioblastoma Cell Culture Platforms Based on Transcriptional Similarity with Paired Tissue.Pharmaceuticals (Basel, Switzerland) · 2024Article
- Establishment of a 96-well transwell system using primary human gut organoids to capture multiple quantitative pathway readouts.Scientific reports · 2023Article
- Advances towards the completeFaculty reviews · 2023Review
Corrections and comments
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Authors and funding
10 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In the expanding field of intestinal organoid research, various protocols for three- and two-dimensional organoid-derived cell cultures exist. Two-dimensional organoid-derived monolayers are used to overcome some limitations of three-dimensional organoid cultures. They are increasingly used also in infection research, to study physiological processes and tissue barrier functions, where easy experimental access of pathogens to the luminal and/or basolateral cell surface is required. This has resulted in an increasing number of publications reporting different protocols and media compositions for organoid manipulation, precluding direct comparisons of research outcomes in some cases. With this in mind, here we describe a protocol aimed at the harmonization of seeding conditions for three-dimensional intestinal organoids of four commonly used research species onto cell culture inserts, to create organoid-derived monolayers that form electrophysiologically tight epithelial barriers. We give an in-depth description of media compositions and culture conditions for creating these monolayers, enabling also the less experienced researchers to obtain reproducible results within a short period of time, and which should simplify the comparison of future studies between labs, but also encourage others to consider these systems as alternative cell culture models in their research. Graphic abstract:
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Registered trials
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.