The DOI matches its Crossref record; 9 of the 9 references it cites were found in a registry or an index.
Isolation and Characterization of Intestinal Stem Cells Based on Surface Marker Combinations and Colony-Formation Assay
Open accessAbstract
BACKGROUND & AIMS Identification of intestinal stem cells (ISCs) has relied heavily on the use of transgenic reporters in mice, but this approach is limited by mosaic expression patterns and difficult to directly apply to human tissues. We sought to identify reliable surface markers of ISCs and establish a robust functional assay to characterize ISCs from mouse and human tissues. METHODS We used immunohistochemistry, real-time reverse-transcription polymerase chain reaction, and fluorescence-activated cell sorting (FACS) to analyze intestinal epithelial cells isolated from mouse and human intestinal tissues. We compared different combinations of surface markers among ISCs isolated based on expression of Lgr5–green fluorescent protein. We developed a culture protocol to facilitate the identification of functional ISCs from mice and then tested the assay with human intestinal crypts and putative ISCs. RESULTS CD44+CD24loCD166+ cells, isolated by FACS from mouse small intestine and colon, expressed high levels of stem cell–associated genes. Transit-amplifying cells and progenitor cells were then excluded based on expression of GRP78 or c-Kit. CD44+CD24loCD166+ GRP78lo/− putative stem cells from mouse small intestine included Lgr5-GFPhi and Lgr5-GFPmed/lo cells. Incubation of these cells with the GSK inhibitor CHIR99021 and the E-cadherin stabilizer Thiazovivin resulted in colony formation by 25% to 30% of single-sorted ISCs. CONCLUSIONS We developed a culture protocol to identify putative ISCs from mouse and human tissues based on cell surface markers. CD44+CD24loCD166+, GRP78lo/−, and c-Kit− facilitated identification of putative stem cells from the mouse small intestine and colon, respectively. CD44+CD24−/loCD166+ also identified putative human ISCs. These findings will facilitate functional studies of mouse and human ISCs.
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Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche
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Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts
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Self-citationMerlos-Suárez, A., Barriga, F. M., Jung, P., Iglesias, M., Céspedes, M. V., Rossell, D., Sevillano, M., Hernando-Momblona, X., da Silva-Diz, V., Muñoz, P., et al. (2011) · Cell Stem Cell · doi:10.1016/j.stem.2011.02.020 · Cited by 934
mTORC1 in the Paneth cell niche couples intestinal stem-cell function to calorie intake
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Clevers, H. (2016) · Cell · doi:10.1016/j.cell.2016.05.082 · Cited by 3,196
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Gjorevski, N., Sachs, N., Manfrin, A., Giger, S., Bragina, M. E., Ordóñez-Morán, P., Clevers, H., & Lutolf, M. P. (2016) · Nature · doi:10.1038/nature20168 · Cited by 1,426
The intestinal crypt, a prototype stem cell compartment
Clevers, H. (2013) · Cell · doi:10.1016/j.cell.2013.07.004 · Cited by 1,223
An in vivo model of human small intestine using pluripotent stem cells
Watson, C. L., Mahe, M. M., Múnera, J., Howell, J. C., Sundaram, N., Poling, H. M., Schweitzer, J. I., Vallance, J. E., Mayhew, C. N., Sun, Y., et al. (2014) · Nature Medicine · doi:10.1038/nm.3737 · Cited by 566
Self-citationVanDussen, K. L., Marinshaw, J. M., Shaikh, N., Miyoshi, H., Moon, C., Tarr, P. I., Ciorba, M. A., & Stappenbeck, T. S. (2015) · Gut · doi:10.1136/gutjnl-2013-306651 · Cited by 525
Metadata
- Authors
- Wang, F., Scoville, D., He, X. C., Mahe, M. M., Box, A., Perry, J. M., Smith, N. R., Lei, N. Y., Davies, P. S., Fuller, M. K., Haug, J. S., McClain, M., Gracz, A. D., Ding, S., Stelzner, M., Dunn, J. C. Y., Magness, S. T., Wong, M. H., Martin, M. G., Helmrath, M., & Li, L.
- Venue
- Gastroenterology
- Publisher
- Elsevier BV
- Published
- 2013-08
- Volume
- 145
- Issue
- 2
- Pages
- 383-395.e21
- ISSN
- 0016-5085 (ISSN-L)
- OpenAlex
- W1991692366
- Cited by
- 219
- References
- 9
- FWCI
- 10.91
- Type
- journal article
- Language
- en
- Open access
- green
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- Wang, F., Scoville, D., He, X. C., Mahe, M. M., Box, A., Perry, J. M., Smith, N. R., Lei, N. Y., Davies, P. S., Fuller, M. K., Haug, J. S., McClain, M., Gracz, A. D., Ding, S., Stelzner, M., Dunn, J. C. Y., Magness, S. T., Wong, M. H., Martin, M. G., . . . Li, L. (2013). Isolation and Characterization of Intestinal Stem Cells Based on Surface Marker Combinations and Colony-Formation Assay. Gastroenterology. https://doi.org/10.1053/j.gastro.2013.04.050
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
