Three-Dimensional Skin Constructs

A method of generating a three-dimensional skin tissue construct comprising a basal-to-suprabasal transition, the method comprising:

• (a) depositing onto a substrate a droplet comprising one or more undifferentiated keratinocyte cells;
• (b) depositing onto the substrate another droplet comprising one or more additional undifferentiated keratinocyte cells;
• and
• (c) maintaining the undifferentiated keratinocyte cells on the substrate in a condition that promotes differentiation of undifferentiated keratinocyte cells;
• wherein the droplet of (b) is deposited a predetermined distance away from the droplet of (a);
• wherein undifferentiated keratinocyte cells migrate from droplets of (a) and (b), generating a basal cell layer;
• wherein proliferation and differentiation of keratinocyte cells of the basal cell layer generate differentiated suprabasal keratinocyte cells and a basal-to-suprabasal transition between at least a subset of adjacent keratinocyte cells of the basal layer and differentiated suprabasal keratinocyte cells;
• and
• wherein the extent of differentiation of keratinocyte cells and the extent of the basal-to suprabasal transition present after (c) varies based on the predetermined distance.