Back to Search
Start Over
Development of Functional Microfold (M) Cells from Intestinal Stem Cells in Primary Human Enteroids
- Source :
- PLoS ONE, PLoS ONE, Vol 11, Iss 1, p e0148216 (2016), PloS one, vol 11, iss 1
- Publication Year :
- 2015
-
Abstract
- Background & Aims Intestinal microfold (M) cells are specialized epithelial cells that act as gatekeepers of luminal antigens in the intestinal tract. They play a critical role in the intestinal mucosal immune response through transport of viruses, bacteria and other particles and antigens across the epithelium to immune cells within Peyer’s patch regions and other mucosal sites. Recent studies in mice have demonstrated that M cells are generated from Lgr5+ intestinal stem cells (ISCs), and that infection with Salmonella enterica serovar Typhimurium increases M cell formation. However, it is not known whether and how these findings apply to primary human small intestinal epithelium propagated in an in vitro setting. Methods Human intestinal crypts were grown as monolayers with growth factors and treated with recombinant RANKL, and assessed for mRNA transcripts, immunofluorescence and uptake of microparticles and S. Typhimurium. Results Functional M cells were generated by short-term culture of freshly isolated human intestinal crypts in a dose- and time-dependent fashion. RANKL stimulation of the monolayer cultures caused dramatic induction of the M cell-specific markers, SPIB, and Glycoprotein-2 (GP2) in a process primed by canonical WNT signaling. Confocal microscopy demonstrated a pseudopod phenotype of GP2-positive M cells that preferentially take up microparticles. Furthermore, infection of the M cell-enriched cultures with the M cell-tropic enteric pathogen, S. Typhimurium, led to preferential association of the bacteria with M cells, particularly at lower inoculum sizes. Larger inocula caused rapid induction of M cells. Conclusions Human intestinal crypts containing ISCs can be cultured and differentiate into an epithelial layer with functional M cells with characteristic morphological and functional properties. This study is the first to demonstrate that M cells can be induced to form from primary human intestinal epithelium, and that S. Typhimurium preferentially infect these cells in an in vitro setting. We anticipate that this model can be used to generate large numbers of M cells for further functional studies of these key cells of intestinal immune induction and their impact on controlling enteric pathogens and the intestinal microbiome.
- Subjects :
- 0301 basic medicine
Salmonella typhimurium
Bacterial Diseases
Confocal Microscopy
Cellular differentiation
Peyers Patches
Cell Culture Techniques
lcsh:Medicine
Small
Pathology and Laboratory Medicine
Epithelium
Peyer's Patches
Intestinal mucosa
Cell Signaling
Stem Cell Research - Nonembryonic - Human
Salmonella
Animal Cells
Intestine, Small
Medicine and Health Sciences
2.1 Biological and endogenous factors
Aetiology
Intestinal Mucosa
lcsh:Science
Cells, Cultured
WNT Signaling Cascade
Microfold cell
Staining
Mucosal
Microscopy
Cultured
Multidisciplinary
Stem Cells
Cell Staining
Light Microscopy
Cell Differentiation
Foodborne Illness
Intestinal epithelium
Signaling Cascades
Intestine
3. Good health
Cell biology
Bacterial Pathogens
Infectious Diseases
Medical Microbiology
Stem cell
Anatomy
Pathogens
Cellular Types
Cellular Structures and Organelles
Research Article
Signal Transduction
General Science & Technology
Cells
Biology
Research and Analysis Methods
Microbiology
Vaccine Related
03 medical and health sciences
Immune system
Antigen
Enterobacteriaceae
Biodefense
Humans
Immunity, Mucosal
Microbial Pathogens
Bacteria
Prevention
lcsh:R
RANK Ligand
Immunity
Organisms
Biology and Life Sciences
Epithelial Cells
Cell Biology
Stem Cell Research
Gastrointestinal Tract
Emerging Infectious Diseases
030104 developmental biology
Biological Tissue
Cell Signaling Structures
Cell culture
Specimen Preparation and Treatment
lcsh:Q
Digestive Diseases
Digestive System
Developmental Biology
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 11
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- PloS one
- Accession number :
- edsair.doi.dedup.....625063d09a52ee1e30c2a3ee14538818