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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Hamilton, Kathryn E. Crissey, Mary Ann S. Lynch, John P. Rustgi, Anil K. |
| Description | Author Affiliation: Hamilton KE ( Gastroenterology Division, Departments of Medicine and Cell and Developmental Biology, Perelman School of Medicine at the University of Pennsylvania and Abramson Cancer Center, Philadelphia, Pennsylvania 19104.); Crissey MA ( Gastroenterology Division, Departments of Medicine and Cell and Developmental Biology, Perelman School of Medicine at the University of Pennsylvania and Abramson Cancer Center, Philadelphia, Pennsylvania 19104.); Lynch JP ( Gastroenterology Division, Departments of Medicine and Cell and Developmental Biology, Perelman School of Medicine at the University of Pennsylvania and Abramson Cancer Center, Philadelphia, Pennsylvania 19104.); Rustgi AK ( Gastroenterology Division, Departments of Medicine and Cell and Developmental Biology, Perelman School of Medicine at the University of Pennsylvania and Abramson Cancer Center, Philadelphia, Pennsylvania 19104.) |
| Abstract | In recent years, the study of primary cells in culture has evolved from an extraphysiological, two-dimensional platform to novel, three-dimensional platforms in which the addition of matrix components and/or supporting cells provide an ex vivo niche. Such studies have provided the basis on which to study more advanced physiological processes in detail, including multilayered, long-term cultures, epithelial-stromal interactions, and stem cell behaviors that more closely recapitulate normal morphology than two-dimensional culture. Various techniques for three-dimensional organotypic culture and crypt culture of primary cells from mouse and human small intestine and colon have been described. These methods have allowed for the study of specific stem cell characteristics, including survival, self-renewal, and long-term growth in culture, as well as the ability to propagate all the appropriate progenitor and postmitotic lineages. These assays have become a widely accepted functional measure of 'stemness' and, in combination with lineage-tracing experiments in various genetically engineered mouse models, have been critical in the identification of specific markers of intestinal stem cells. In this protocol we draw upon recently described methods for the isolation and culture of mouse small intestinal enterospheres/enteroids from isolated crypts and/or single cells. Cultures of murine colon epithelium, as well as human small intestine and colon, require additional growth factors not discussed here. The description provided here represents current knowledge, and it is possible, if not likely, that modifications in the future will emerge. |
| File Format | HTM / HTML |
| ISSN | 19403402 |
| Issue Number | 4 |
| Volume Number | 2015 |
| e-ISSN | 15596095 |
| Journal | Cold Spring Harbor Protocols |
| Language | English |
| Publisher | Cold Spring Harbor Laboratory Press |
| Publisher Date | 2015-04-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Clinical Laboratory Techniques Adult Stem Cells Cytology Cell Culture Techniques Methods Intestine, Small Animals Cell Separation Cells, Cultured Mice Journal Article Research Support, N.i.h., Extramural |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry, Genetics and Molecular Biology |
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