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Temporal-spatial protein expression in bladder tissue derived from embryonic stem cells.
Thomas JC, Oottamasathien S, Makari JH, Honea L, Sharif-Afshar AR, Wang Y, Adams C, Wills ML, Bhowmick NA, Adams MC, Brock JW, Hayward SW, Matusik RJ, Pope JC
(2008) J Urol 180: 1784-9
MeSH Terms: Animals, Embryonic Stem Cells, Female, Gene Expression Regulation, Developmental, Hepatocyte Nuclear Factor 3-alpha, Hepatocyte Nuclear Factor 3-beta, Immunohistochemistry, Male, Membrane Glycoproteins, Mice, Mice, Nude, Pregnancy, Rats, Rats, Sprague-Dawley, Stem Cells, Tissue Engineering, Transplantation, Heterologous, Urinary Bladder, Uroplakin III
Show Abstract · Added November 28, 2012
PURPOSE - Identifying developmental proteins could lead to markers of bladder progenitor cells, which could be used to investigate bladder diseases. We recently reported a novel embryonic stem cell model in which to study differential protein expression patterns during bladder development. Differential and temporal expressions of the endodermal proteins known as forkhead box (Foxa1 and Foxa2) were observed. In the current study we further delineated these protein expression patterns.
MATERIALS AND METHODS - Epithelium was removed from the underlying mesenchyma from embryonic day 18 rat bladders. Heterospecific recombinant xenografts were created by combining embryonic stem cells plus embryonic bladder mesenchyma and placed beneath the renal capsule of mouse hosts. Grafts were harvested at 16, 18, 21, 28, 35 and 42 days, and evaluated with hematoxylin and eosin, trichrome staining, and immunohistochemistry for uroplakin, smooth muscle alpha-actin, p63, Foxa1, Foxa2 and androgen receptor.
RESULTS - At 16 days uroplakin was detectable and it seemed to correlate with the loss of Foxa2, while Foxa1 remained at all time points. Androgen receptor was first noted in stroma at day 16. It localized to urothelial nuclei at day 21 and was undetectable at 42 days. Adjacent to the urothelium alpha-smooth muscle actin was seen on day 16 and it was localized in bundles to the periphery of the graft at later time points. Staining for basilar urothelium with p63 confirmed basilar orientation at all time points.
CONCLUSIONS - We report the temporal spatial expression of various genes in early bladder development. This suggests that some proteins may be potential markers of bladder progenitor cells. Characterizing these markers may potentially identify bladder progenitor cells that have been directed toward a lineage path destined to become urothelial cells. Ultimately these multipotential progenitor cells could be isolated and used to study and treat diseases that affect the bladder.
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19 MeSH Terms
Bladder tissue formation from cultured bladder urothelium.
Oottamasathien S, Williams K, Franco OE, Thomas JC, Saba K, Bhowmick NA, Staack A, Demarco RT, Brock JW, Hayward SW, Pope JC
(2006) Dev Dyn 235: 2795-801
MeSH Terms: Animals, Blotting, Western, Cell Differentiation, Cells, Cultured, Female, Immunohistochemistry, Male, Membrane Glycoproteins, Mesoderm, Mice, Mice, Nude, Pregnancy, Rats, Rats, Sprague-Dawley, Tissue Engineering, Transplantation, Heterologous, Urinary Bladder, Uroplakin III, Urothelium
Show Abstract · Added December 10, 2013
Tissue recombination is a powerful method to evaluate the paracrine-signaling events that orchestrate the development of organs using the in vivo environment of a host rodent. Studies have reported the successful generation of primary cultures of rodent bladder urothelium, but none have reported their use to recapitulate bladder tissue with tissue recombination. We propose that primary cultured bladder urothelium, when recombined with inductive embryonic bladder mesenchyme, will form bladder tissue in a recombination model. Adult rat bladders were isolated and urothelium obtained. Sheets of bladder urothelium were re-suspended in collagen and maintained in tissue culture. After expansion (>20 passages), the urothelium was recombined with embryonic day-14 mouse bladder mesenchyme, then grafted beneath the renal capsule of immunocompromised mouse hosts. Grafts were harvested after 28 days. Control grafts were performed with bladder mesenchyme alone, cultured bladder urothelium alone, and collagen matrix alone. Final tissues were evaluated with staining and immunohistochemistry (H&E, Gomori's trichrome, broad-spectrum uroplakin, and smooth muscle actin alpha and gamma). Immunocytochemistry on cultured urothelium for broad-spectrum keratin, vimentin, and broad-spectrum uroplakin confirmed pure populations, void of mesenchymal contaminants. Staining of recombinant grafts demonstrated bladder tissue with mature urothelium and stromal differentiation. Control tissues were void of bladder tissue formation. We have successfully demonstrated that a chimeric bladder is formed from primary cultured bladder urothelium recombined with embryonic bladder mesenchyme. This is a powerful new tool for investigating the molecular mechanisms of bladder development and disease. Future applications may include the in vitro genetic manipulation of urothelium and examining those effects on growth and development in an in vivo environment.
(c) 2006 Wiley-Liss, Inc.
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1 Members
0 Resources
19 MeSH Terms