Validation and quantitation of an in vitro M-cell model

• Published in: Biochemical and biophysical research communications 2002-12, Vol.299 (3), p.377-383
• Main Authors: Tyrer, Peter, Ruth Foxwell, A, Kyd, Jennelle, Harvey, Matthew, Sizer, Phillip, Cripps, Allan
• Format: Article
• Language: English
• Subjects: Alkaline Phosphatase - metabolism; Animals; Caco-2; Caco-2 Cells; Carrier Proteins - metabolism; Cell Culture Techniques - methods; Cells, Cultured; Coculture Techniques; Follicle-associated epithelium; Humans; Integrin alpha5beta1 - metabolism; Intestinal Mucosa - cytology; Intestinal Mucosa - metabolism; Lymphocyte Subsets - cytology; Lymphocyte Subsets - metabolism; M-cell; Mice; Mice, Inbred BALB C; Microfilament Proteins - metabolism; Microspheres; Models, Biological; Peyer's Patches - cytology; Peyer’s patch; Reproducibility of Results; Vaccine
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/S0006-291X(02)02631-1

This study has evaluated an in vitro model of the follicle-associated epithelia that overlie Peyer’s patches of the small intestine. The model shares many phenotypic characteristics of M cells in vivo. Co-cultures of the human adenocarcinoma cell line Caco-2 and freshly isolated Peyer’s patch cells were established. Fluorescence microscopy and quantitative image analysis were used to validate the model against known markers of M-cell phenotype. Apical expression of alkaline phosphatase was down-regulated in co-cultures and villin was re-distributed from the apical membrane to the cytoplasm. α5β1 integrin was found on the apical surfaces of the monolayers and B and T lymphocytes integrated into the monolayers. Particle transport was temperature-dependent in co-cultures, indicating that a transcytotic route was responsible. This model provides opportunities to study factors that influence M-cell development, assess putative Peyer’s patch targeting in oral vaccine technologies, and study intestinal uptake in vitro.