Regenerative Medicine (2003) / Chapter Skim
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14 In vivo regeneration of murine prostate from dissocated cell populations of postnatal epithelia and urogenital sinus mesenchyme
14 In vivo regeneration of murine prostate from dissocated cell populations of postnatal epithelia and urogenital sinus mesenchyme
Pages 80-87
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From page 80... ...
A variety of evidence from immunohistochemical analyses monitoring differential expression of cytokeratins and other markers suggests that transitional cell types between the basal and luminal types can be defined as important intermediates in this developmental sequence (17-19~. Prostatic stem cells in the adult mouse or rat can be defined by their ability to survive acute androgen withdrawal and to differentiate and reform a prostate on androgen add back (18~.
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From page 81... ...
Our long-term goal is to define in a physical and functional manner the subpopulation of cells in the rodent prostate with the essential characteristics of stem cells. We needed a system in which dispersed cell populations rather than tissue fragments could be analyzed to eventually define surface antigens and other markers to fractionate the population, enrich for cells capable of tissue structure reconstitution, develop appropriate methods for clonally marking individual cells, and genetically manipulate the epithelial and mesenchymal populations.
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From page 82... ...
RNA Expression Analysis. Total RNAs from the adult mouse prostate, seminal vesicle, urethra, bladder, liver, and regenerated prostate were extracted with an RNeasy Mini kit (Qiagen, Valencia, CA)
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From page 83... ...
Results Single-Cell Populations of UGSM and Postnatal Prostate Epithelial Cells Can Combine to Regenerate Prostatic-Like Structures in vivo. Self-renewal, clonogenicity, and tissue regeneration are essential characteristics of stem cells.
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From page 84... ...
Discussion In this work, we have defined the experimental conditions to regenerate prostate tissue from dissociated cell populations of UGS-derived mesenchyme and prostate epithelial populations taken from postnatal animals from 10 d to 6 wk of age. These single-cell suspensions allow for accurate quantitation, uniform introduction of visual markers like GFP by viral vector technologies, and eventually selectable markers or genes, which may influence the growth and developmental phenotype of the regenerated tissue.
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From page 85... ...
UGSM cells (1 x 105) were mixed with 10-d-old p-actin GFP mouse prostate epithelial cells ranging from 5 x 103 to 5 x 104, and the regeneration protocol was carried out as described in Materials and Methods and Fig.
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From page 86... ...
Limiting dilution colony assays on specific feeder cell lines for hematopoietic stem cells to produce cells of myeloid and lymphoid lineages has been particularly useful in this setting (56~. Several groups have reported such short-term colony-like assays for prostate epithelial cells, and these should be evaluated for their ability to enumerate or even expand cells with regenerative activity in vivo (26-28~.
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From page 87... ...
Xin et al. and Gerry Cunha and members of the Cunha laboratory for introducing us to the surgical techniques essential for this study, the area of prostate regeneration by tissue transplantation, and the polyclonal antiserum anti-secretions of mouse dorsolateral prostate.
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