Maintenance of functional stem cells in isolated and cultured adult intestinal epithelium.

2.50
Hdl Handle:
http://hdl.handle.net/10541/90796
Title:
Maintenance of functional stem cells in isolated and cultured adult intestinal epithelium.
Authors:
Booth, Catherine; O'Shea, Julie A; Potten, Christopher S
Abstract:
We have previously described a method for the primary culture of adult large intestinal epithelium, suggesting that stem cells had survived both the isolation and the culture procedures. However, as no markers for such cells exist, confirmation of stem cell survival is difficult-only the functional properties can be used to define them. Unfortunately, many of these (e.g., differentiation, crypt regeneration) do not occur in culture, probably due to suboptimal conditions. To address this problem both freshly isolated and cultured small and large intestinal crypts were grown subcutaneously in an immunocompromized mouse. All initially formed cysts lined by a simple epithelium which gradually became multicellular and formed invaginations containing many mitoses and apoptoses. Epithelial differentiation, as assayed by Goblet cell mucin production, was also apparent. Mucin maturation was also typical of the normal intestine. The lumen was frequently filled with mucin and apoptotic bodies. Interestingly, in grafts displaying pronounced crypt-like morphology the regions of proliferation were situated toward the base of the structure and the Goblet cells toward the lumen, i.e., a typical crypt-like morphology. Hence, functional adult stem cells appear to survive isolation and tissue culture, permitting organotypic regeneration, possibly involving homeobox gene expression. This may now allow direct stem cell characterization and experimental manipulation, such as transfection, and may ultimately permit transplantation and therapeutic gene therapy.
Affiliation:
Epithelial Biology Group, CRC Section of Cell and Tumour Biology, Paterson Institute, Christie Hospital (NHS) Trust, Manchester, M20 4BX, United Kingdom. CBooth@picr.man.ac.uk
Citation:
Maintenance of functional stem cells in isolated and cultured adult intestinal epithelium. 1999, 249 (2):359-66 Exp. Cell Res.
Journal:
Experimental Cell Research
Issue Date:
15-Jun-1999
URI:
http://hdl.handle.net/10541/90796
DOI:
10.1006/excr.1999.4483
PubMed ID:
10366435
Type:
Article
Language:
en
ISSN:
0014-4827
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorBooth, Catherineen
dc.contributor.authorO'Shea, Julie Aen
dc.contributor.authorPotten, Christopher Sen
dc.date.accessioned2010-01-28T11:40:50Z-
dc.date.available2010-01-28T11:40:50Z-
dc.date.issued1999-06-15-
dc.identifier.citationMaintenance of functional stem cells in isolated and cultured adult intestinal epithelium. 1999, 249 (2):359-66 Exp. Cell Res.en
dc.identifier.issn0014-4827-
dc.identifier.pmid10366435-
dc.identifier.doi10.1006/excr.1999.4483-
dc.identifier.urihttp://hdl.handle.net/10541/90796-
dc.description.abstractWe have previously described a method for the primary culture of adult large intestinal epithelium, suggesting that stem cells had survived both the isolation and the culture procedures. However, as no markers for such cells exist, confirmation of stem cell survival is difficult-only the functional properties can be used to define them. Unfortunately, many of these (e.g., differentiation, crypt regeneration) do not occur in culture, probably due to suboptimal conditions. To address this problem both freshly isolated and cultured small and large intestinal crypts were grown subcutaneously in an immunocompromized mouse. All initially formed cysts lined by a simple epithelium which gradually became multicellular and formed invaginations containing many mitoses and apoptoses. Epithelial differentiation, as assayed by Goblet cell mucin production, was also apparent. Mucin maturation was also typical of the normal intestine. The lumen was frequently filled with mucin and apoptotic bodies. Interestingly, in grafts displaying pronounced crypt-like morphology the regions of proliferation were situated toward the base of the structure and the Goblet cells toward the lumen, i.e., a typical crypt-like morphology. Hence, functional adult stem cells appear to survive isolation and tissue culture, permitting organotypic regeneration, possibly involving homeobox gene expression. This may now allow direct stem cell characterization and experimental manipulation, such as transfection, and may ultimately permit transplantation and therapeutic gene therapy.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshCells, Cultured-
dc.subject.meshCrosses, Genetic-
dc.subject.meshFemale-
dc.subject.meshIntestinal Mucosa-
dc.subject.meshMale-
dc.subject.meshMice-
dc.subject.meshMice, Inbred NOD-
dc.subject.meshMice, SCID-
dc.subject.meshStem Cell Transplantation-
dc.subject.meshStem Cells-
dc.titleMaintenance of functional stem cells in isolated and cultured adult intestinal epithelium.en
dc.typeArticleen
dc.contributor.departmentEpithelial Biology Group, CRC Section of Cell and Tumour Biology, Paterson Institute, Christie Hospital (NHS) Trust, Manchester, M20 4BX, United Kingdom. CBooth@picr.man.ac.uken
dc.identifier.journalExperimental Cell Researchen

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