Figure 6

Effect of essential fatty acids on ePUKs morphology, ePUKs on microbeads, and growth on EVPOME. 6a-c: Essential fatty acids and palmitic acid (16:0) were prepared and added to the medium: 5 μM 16:0, and 10 μM 18:2 (n-6) and 20:4 (n-6) bound to bovine serum albumin as carrier. a: control T = 0, b = 24 hr. and c = 48 hr. culture. Bar = 50 microns. 6d: The ePUK cells can be transferred to microcarrier beads. Bar = 50 microns. 6e and f: Mucosal cells grown from the retromoral pad were placed on Alloderm®, air lifted for 11 days and processed. Stain was Hematoxylin/Eosin, arrows denote basal layer and the stratified differentiated layers stain pink. e = ePUK cells and f = monolayer from which the cells were derived. The ePUKS were at P1S1 as defined in Figure 2a.The is Bar = 100 microns. 6 g and h: adult human epidermis derived ePUKs, P1S1p3 (Figure 2a), formed a stratified multilayered structure on Alloderm®, day 11; stained with H&E (Figure 6g). The structure stained for k10 keratins (Figure 6h), specific for epithelial differentiation in the uppermost layers (arrow). Bar is 100 microns. 6i: adult human epidermis derived ePUKs formed an EVPOME using P1S1p3 cells as defined in Figure 2a. Figure 6 i = an EVPOME stained with anti-filaggrin antibody showing production of this epidermal differentiation-specific protein by the upper most layer formed by ePUK derived cells (arrow). Bar = 100 microns.