Validation and quantitation of an in vitro M-cell model.
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. alpha5beta1 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.
Tyrer
Peter
P
Gadi Research Centre, Division of Science and Design, University of Canberra, ACT 2601, Canberra, Australia.
Ruth Foxwell
A
A
Kyd
Jennelle
J
Harvey
Matthew
M
Sizer
Phillip
P
Cripps
Allan
A
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D018938:Caco-2 Cells; D002352:Carrier Proteins / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D018920:Coculture Techniques; D006801:Humans; D039081:Integrin alpha5beta1 / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology* / Q000378:metabolism*; D016131:Lymphocyte Subsets / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008840:Microfilament Proteins / Q000378:metabolism; D008863:Microspheres; D008954:Models, Biological; D010581:Peyer's Patches / Q000166:cytology*; D015203:Reproducibility of Results
D002352:Carrier Proteins; D039081:Integrin alpha5beta1; D008840:Microfilament Proteins; C026615:villin; D000469:Alkaline Phosphatase