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Updated: Feb 25, 2026
Application of a Mouse Ligated Peyer’s Patch Intestinal Loop Assay to Evaluate Bacterial Uptake by M cells
Published on: December 17, 2011
A comparison of three Peyer's patch "M-like" cell culture models: particle uptake, bacterial interaction, and
Tauseef Ahmad1, Martina Gogarty1, Edwin G Walsh1
1UCD School of Veterinary Medicine and UCD Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland.
Abstract:
Intestinal Peyer's patch (PP) microfold (M) cells transport microbes and particulates across the follicle-associated epithelium (FAE) as part of the mucosal immune surveillance system. In vitro human M-like cell co-culture models are used as screens to investigate uptake of antigens-in-nanoparticles, but the models are labour-intensive and there is inter-laboratory variability. We compared the three most established filter-grown Caco-2/Raji B cell co-culture systems. These were Model A (Kernéis et al., 1997), Model B (Gullberg et al., 2000), and Model C (Des Rieux et al. 2007). The criteria used were transepithelial resistance (TEER), the apparent permeability coefficient (Papp) of [14C]-mannitol, M cell-like histology, as well as latex particle and Salmonella typhimurium translocation. Each co-culture model displayed substantial increases in particle translocation. Truncated microvilli compared to mono-cultures was their most consistent feature. The inverted model developed by des Rieux et al. (2007) displayed reductions in TEER and an increased (Papp), accompanied by the largest increase in particle translocation compared to the other two models. The normally-oriented model developed by Gullberg et al. (2000) was the only one to consistently display an increased translocation of Salmonella typhimurium. By applying a double Matrigel™ coating on filters, altering the medium feeding regime for Raji B cells, and restricting the passage number of B cells, improvements to the Gullberg model B were achieved, as reflected by increased particle translocation and improved histology. In conclusion, this is the first time all three designs have been compared in one study and each displays phenotypic features of M-like cells. While Model C was the most robust co-culture, the Model B protocol could be improved by optimizing several variables and is less complicated to establish than the two inverted models.
Insights
This study compares three in vitro models of M-like cells for studying mucosal immunity. Model C showed the most robust particle translocation, while Model B could be optimized for improved performance.
Area of Science:
- Immunology
- Cell Biology
- Gastroenterology
Background:
- Intestinal Peyer's patch (PP) microfold (M) cells are crucial for mucosal immune surveillance, transporting microbes and particulates.
- In vitro M-like cell co-culture models are valuable for studying antigen uptake but suffer from labor intensity and variability.
- Comparing established Caco-2/Raji B cell co-culture systems is essential for optimizing these models.
Purpose of the Study:
- To compare three established filter-grown Caco-2/Raji B cell co-culture systems (Models A, B, and C) used to mimic M cell function.
- To evaluate these models based on transepithelial electrical resistance (TEER), permeability, histology, and translocation of particles and bacteria.
- To identify the most robust and potentially improvable model for M cell research.
Main Methods:
- Comparison of three filter-grown Caco-2/Raji B cell co-culture systems: Model A (Kernéis et al., 1997), Model B (Gullberg et al., 2000), and Model C (Des Rieux et al., 2007).
- Assessment of transepithelial electrical resistance (TEER) and apparent permeability coefficient (Papp) using [14C]-mannitol.
- Evaluation of M cell-like histology, latex particle translocation, and Salmonella Typhimurium translocation.
Main Results:
- All three co-culture models demonstrated increased particle translocation and truncated microvilli compared to mono-cultures.
- Model C (inverted) showed reduced TEER, increased Papp, and the highest particle translocation.
- Model B (normally-oriented) was unique in consistently showing increased Salmonella Typhimurium translocation; improvements were achieved via double Matrigel coating and optimized Raji B cell culture.
- Model C proved the most robust, while Model B offered a less complex, improvable alternative.
Conclusions:
- All three models exhibit M-like cell phenotypic features, validating their use in research.
- Model C is the most robust co-culture system for M cell research.
- Model B, while less complex, can be significantly improved through protocol optimization, offering a viable alternative.
