PMC

Native adult mouse bladder. a. Immunohistochemical detection of broad-spectrum uroplakin (brown) denoting mature urothelium. b. Immunohistochemical detection of p63 (brown) demonstrating urothelial basilar orientation. c. Immunohistochemical detection of Foxa1 (brown) seen in all urothelial cells denoting endodermal derivation of the cells. d. Complete lack of immunohistochemical detection of Foxa2.

Native postnatal day 1 of life mouse bladder. a. Immunohistochemical detection of broad-spectrum uroplakin (brown) denoting mature urothelium. b. Immunohistochemical detection of p63 (brown) demonstrating urothelial basilar orientation. c. Immunohistochemical detection of Foxa1 (brown) seen in all urothelial cells denoting endodermal derivation of the cells. d. Complete lack of immunohistochemical detection of Foxa2.

Xenografts of 1500 embryonic stem cells + 4 embryonic bladder mesenchymal shells/graft harvested at 42 days. a&b. Gomori’s trichrome, multilayered cuboidal urothelial cells (red) with sub-urothelial connective tissue (blue) were seen. c&d. Immunohistochemical detection of smooth muscle α-actin (brown) denoting smooth muscle fibrils. e&f. Immunohistochemical detection of broad-spectrum uroplakin (brown) denoting mature urothelium.

Native embryonic day 16 mouse bladder. a. Immunohistochemical detection of broad-spectrum uroplakin (brown) denoting mature urothelium. b. Immunohistochemical detection of p63 (brown) demonstrating urothelial basilar orientation. c. Immunohistochemical detection of Foxa1 (brown) seen in all urothelial cells denoting endodermal derivation of the cells. d. Immunohistochemical detection of Foxa2 (brown), arrow denotes one positive staining cell within the section.

Xenografts of 1500 embryonic stem cells + 4 embryonic bladder mesenchymal shells/graft harvested at 7 (a,c,e,g) and 14 (b,d,f,h) days. a&b. Gomori’s trichrome, cellular organization occurring in cells that look destined to become urothelium (blue). c&d. Immunohistochemical detection of p63 (brown) illustrating lack of expression within 7 day tissues and early expression observed in 14 day tissues. e&f. Immunohistochemical detection of Foxa1 (brown) denoting endodermal cells. g&h. Immunohistochemical detection of Foxa2 (brown) denoting endodermal cells.

Xenografts harvested at 42 days. a. Gross appearance of 1000 embryonic stem (ES) cell grafts on host kidney (arrowheads denote whitish-tan appearing grafts). b–d. Histology and immunohistochemical staining of 1000 ES cell + 1 embryonic bladder mesenchyme shell recombinant graft. b. Gomori’s trichrome, star denotes central lumen of bladder structure, urothelial cells surrounding central lumen (red) with sub-urothelial connective tissue (blue) were seen. c. Immunohistochemical detection of broad-spectrum uroplakin (brown) denoting mature urothelium. d. Immunohistochemical detection of smooth muscle α-actin (brown) denoting smooth muscle fibrils.

Xenografts of 1500 embryonic stem (ES) cells + 4 embryonic bladder mesenchymal shells/graft harvested at 42 days. a. H&E staining. b. Immunohistochemical detection of p63 (brown) illustrating complete spatial orientation of ES cell derived urothelium. c. Immunohistochemical detection of Foxa1 (brown) seen in all ES cell derived urothelium. d. Immunohistochemical detection of Foxa2 (brown) seen strongly positive in only a few urothelial cells. e&f. H&E and immunohistochemical detection of neuronal nuclear antigen A60 (NeuN) (brown) confirming the presence of neuronal cells clustering in the form of a ganglion. Scale bars of 20 μm apply to images a–d. Scale bars of 10 μm apply to images e&f.