FIG. 2.

Before using Alloderm^®, in preliminary experiments, oral (*) and skin keratinocytes (+) were seeded in Petri dishes with a PDMS barrier (black line in upper right quadrant) to determine whether they will mingle and form the equivalent of a vermillion. (A) Initial seeding, (B) oral and skin keratinocytes advancing fronts “mix” in the middle to form a “vermillion” after migrating in the center of the Petri dish, and (C) fixed and stained oral and skin keratinocytes after 6 days in culture showing confluence and “mixing” of two cell populations. We experimented with barriers of varied thicknesses and lengths to separate the areas where the cells would be seeded to adjust to the sizes of Petri dishes (D, left) or six-well cultures (D, right pictured without the barrier with the central area indicated with black lines). The process is explained in the sequence of image D. The plastic barrier is filled with PDMS in a separate container to allow curing of the PDMS and then used by placing it to adjust over the Alloderm^® (tissue at the bottom in center D) and the container. The barrier shown in D works as a mold to pour the PDMS but several variations of this technique can be implemented to fit the needs of the researcher or to adjust to clinical applications. Color images available online at www.liebertonline.com/tec