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Intestinal subepithelial myofibroblasts support in vitro and in vivo growth of human small intestinal epithelium
- Source :
- PLoS ONE, Vol 6, Iss 11, p e26898 (2011), PloS one, vol 6, iss 11, PLoS ONE
- Publication Year :
- 2011
- Publisher :
- Public Library of Science (PLoS), 2011.
-
Abstract
- The intestinal crypt-niche interaction is thought to be essential to the function, maintenance, and proliferation of progenitor stem cells found at the bases of intestinal crypts. These stem cells are constantly renewing the intestinal epithelium by sending differentiated cells from the base of the crypts of Lieberkühn to the villus tips where they slough off into the intestinal lumen. The intestinal niche consists of various cell types, extracellular matrix, and growth factors and surrounds the intestinal progenitor cells. There have recently been advances in the understanding of the interactions that regulate the behavior of the intestinal epithelium and there is great interest in methods for isolating and expanding viable intestinal epithelium. However, there is no method to maintain primary human small intestinal epithelium in culture over a prolonged period of time. Similarly no method has been published that describes isolation and support of human intestinal epithelium in an in vivo model. We describe a technique to isolate and maintain human small intestinal epithelium in vitro from surgical specimens. We also describe a novel method to maintain human intestinal epithelium subcutaneously in a mouse model for a prolonged period of time. Our methods require various growth factors and the intimate interaction between intestinal sub-epithelial myofibroblasts (ISEMFs) and the intestinal epithelial cells to support the epithelial in vitro and in vivo growth. Absence of these myofibroblasts precluded successful maintenance of epithelial cell formation and proliferation beyond just a few days, even in the presence of supportive growth factors. We believe that the methods described here can be used to explore the molecular basis of human intestinal stem cell support, maintenance, and growth.
- Subjects :
- Pathology
Anatomy and Physiology
Time Factors
Mouse
Cellular differentiation
Digestive Physiology
Cell Culture Techniques
Gene Expression
lcsh:Medicine
Mice, SCID
Regenerative Medicine
Small
Inbred C57BL
Oral and gastrointestinal
Extracellular matrix
Mice
Engineering
Stem Cell Research - Nonembryonic - Human
Mice, Inbred NOD
Molecular Cell Biology
Intestine, Small
2.1 Biological and endogenous factors
Aetiology
Stem Cell Niche
Intestinal Mucosa
Myofibroblasts
lcsh:Science
Cells, Cultured
Mice, Knockout
Cultured
Multidisciplinary
Reverse Transcriptase Polymerase Chain Reaction
Stem Cells
Animal Models
Immunohistochemistry
Intestinal epithelium
Intestine
Cell biology
Adult Stem Cells
medicine.anatomical_structure
Medicine
Muscle
Intercellular Signaling Peptides and Proteins
Stem Cell Research - Nonembryonic - Non-Human
Smooth
Cellular Types
Stem cell
Myofibroblast
Research Article
Biotechnology
Cell Physiology
medicine.medical_specialty
Cell type
General Science & Technology
1.1 Normal biological development and functioning
Cells
Knockout
Biomedical Engineering
Bioengineering
Gastroenterology and Hepatology
Biology
SCID
Model Organisms
Underpinning research
medicine
Animals
Humans
Vimentin
Progenitor cell
Cell Proliferation
Digestive Regulation
Tissue Engineering
lcsh:R
Muscle, Smooth
Epithelial Cells
Stem Cell Research
Actins
Coculture Techniques
Epithelium
Mice, Inbred C57BL
Enterocytes
Inbred NOD
lcsh:Q
Digestive Diseases
Digestive System
Developmental Biology
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 6
- Issue :
- 11
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....1b61d7bda28f94df5b3da434b63539be