ArticleBiological procedures online2020
Caco-2 Cells for Measuring Intestinal Cholesterol Transport - Possibilities and Limitations.
Article in Biological procedures online, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Co-exposure to polystyrene nanoparticles and tetrabromobisphenol A induces genotoxicity and interferes with intestinal barrier differentiation.Cell biology and toxicology · 2026Article
- In vitro evaluation of the anthelmintic activity of citrus flavonoids against free-living and parasitic nematodes.Scientific reports · 2025Article
- Pinoresinol enhances intestinal tight junction integrity via activation of the CaMKKβ-AMPKα1 pathway in Caco-2 cells.Genes & nutrition · 2025Article
- Human Small Intestinal Tissue Models to Assess Barrier Permeability: Comparative Analysis of Caco-2 Cells, Jejunal and Duodenal Enteroid-Derived Cells, and EpiIntestinalBioengineering (Basel, Switzerland) · 2025Article
- Advancements and Challenges in Modeling Mechanobiology in Intestinal Host-Microbiota Interaction.ACS applied materials & interfaces · 2025Review
- Honokiol, a Neolignan from Magnolia officinalis, Attenuated Fructose-Induced Hepatic Fat Accumulation by Improving Intestinal Barrier Function in Mice.The Journal of nutrition · 2025Article
- Comparative analysis of Caco-2 cells and human jejunal and duodenal enteroid-derived cells in gel- and membrane-based barrier models of intestinal permeability.Toxicological sciences : an official journal of the Society of Toxicology · 2025Article
- Article
- Essential amino acids preserve intestinal barrier integrityFrontiers in pharmacology · 2025Article
- Reprogramming human urine cells into intestinal organoids with long-term expansion ability and barrier function.Heliyon · 2024Article
- Improving the Accuracy of Permeability Data to Gain Predictive Power: Assessing Sources of Variability in Assays Using Cell Monolayers.Membranes · 2024Review
- Whey Protein Sodium-Caseinate as a Deliverable Vector for EGCG: In Vitro Optimization of Its Bioaccessibility, Bioavailability, and Bioactivity Mode of Actions.Molecules (Basel, Switzerland) · 2024Article
- Enhanced intestinal epithelial co-culture model with orbital mechanical stimulation: a proof-of-concept application in food nanotoxicology.Frontiers in molecular biosciences · 2024Article
- Caco-2 Cell Line Standardization with Pharmaceutical Requirements and In Vitro Model Suitability for Permeability Assays.Pharmaceutics · 2023Review
- Ion channels and transporters regulate nutrient absorption in health and disease.Journal of cellular and molecular medicine · 2023Review
- Dietary fat induced chylomicron-mediated LPS translocation in a bicameral Caco-2cell model.Lipids in health and disease · 2023Article
- Article
- Intestinal ischemia-reperfusion induces the release of IL-17A to regulate cell inflammation, apoptosis and barrier damage.Experimental and therapeutic medicine · 2022Article
- Article
- Hydroxycinnamic acid derivatives effect on hypercholesterolemia, comparison with ezetimibe: Permeability assays and FTIR spectroscopy on Caco-2 cell line.Current research in pharmacology and drug discovery · 2022Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe human Caco-2 cell line is a common in vitro model of the intestinal epithelial barrier. As the intestine is a major interface in cholesterol turnover and represents a non-biliary pathway for cholesterol excretion, Caco-2 cells are also a valuable model for studying cholesterol homeostasis, including cholesterol uptake and efflux. Currently available protocols are, however, either sketchy or not consistent among different laboratories. Our aim was therefore to generate a collection of optimized protocols, considering the different approaches of the different laboratories and to highlight possibilities and limitations of measuring cholesterol transport with this cell line.
resultsWe developed comprehensive and quality-controlled protocols for the cultivation of Caco-2 cells on filter inserts in a single tight monolayer. A cholesterol uptake as well as a cholesterol efflux assay is described in detail, including suitable positive controls. We further show that Caco-2 cells can be efficiently transfected for luciferase reporter gene assays in order to determine nuclear receptor activation, main transcriptional regulators of cholesterol transporters (ABCA1, ABCB1, ABCG5/8, NPC1L1). Detection of protein and mRNA levels of cholesterol transporters in cells grown on filter inserts can pose challenges for which we highlight essential steps and alternative approaches for consideration. A protocol for viability assays with cells differentiated on filter inserts is provided for the first time.
conclusionsThe Caco-2 cell line is widely used in the scientific community as model for the intestinal epithelium, although with highly divergent protocols. The herein provided information and protocols can be a common basis for researchers intending to use Caco-2 cells in the context of cellular cholesterol homeostasis.
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