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

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    Authors
    Booth, Catherine
    O'Shea, Julie A
    Potten, Christopher S
    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
    Issue Date
    1999-06-15
    
    Metadata
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    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.
    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
    URI
    http://hdl.handle.net/10541/90796
    DOI
    10.1006/excr.1999.4483
    PubMed ID
    10366435
    Type
    Article
    Language
    en
    ISSN
    0014-4827
    ae974a485f413a2113503eed53cd6c53
    10.1006/excr.1999.4483
    Scopus Count
    Collections
    All Paterson Institute for Cancer Research

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