FIG. 1.

Preparation of the amniotic membrane (AM) scaffolds. (A–C) AMs were thawed, washed with sterile phosphate-buffered saline and culture medium, and then fastened with the membrane holders of 14 or 25 mm in diameter. (D–F) Schematic representation of three different AM scaffolds. Urothelial cells (UCs) were seeded on (D, D′, D′′) epithelial AM (eAM), which is directly on amniotic epithelial cells (AEC), on (E, E′, E′′) denuded AM (dAM), which is on amniotic basal lamina (BL), and on (F, F′, F′′) stromal AM (sAM), which is on the spongy layer (SL) of AM stroma (AMS). Before UC seeding, the histology and topography of each AM scaffold was evaluated. Histological sections showed uniform lining of AEC on eAM and sAM scaffolds (D′, F′) and completely removed AECs after thermolysin treatment (E′). Scanning electron micrographs showed (D′′) the apical surface of AECs, (E′′) amniotic basal lamina after thermolysin treatment, and (F′′) spongy layer of sAM. Scale bars: (D′–F′) 100 μm, (D′′–F′′) 10 μm. Color images available online at www.liebertpub.com/tec