Biological and physical factors influencing the successful engraftment of a cultured human skin substitute.

Skin tissue may be engineered in a variety of ways. Our cultured skin substitute (Graftskin, living skin equivalent or G-LSE), Apligraftrade mark, is an organotypic culture of skin, containing both a "dermis" and "epidermis." The epidermis is an important functional component of skin, responsible for biologic wound closure.

Development of a stratum corneum and barrier function in an organotypic skin culture.

The stratum corneum of human skin is responsible for maintaining the epidermal permeability barrier. We have developed a bilayered skin culture (SC) which forms a corneum 35 +/- 1 cell layers thick 21 days after being raised to the air-liquid (A/L) interface. By the 7th day after raising to the A/L interface the corneocytes were irregularly shaped and had cross-sectional areas (CSA) of > or = 300 microns 2.

The organotypic culture of human skin keratinocytes and fibroblasts to achieve form and function.

We describe an organotypic model of human skin comprised of a stratified layer of human epidermal keratinocytes and dermal fibroblasts within a contracted collagen lattice. Feasible and reproducible production of the skin construct has required the use of traditional as well as specialized culture techniques. The configuration of the construct has been engineered to maintain polarity and permit extended culture at the air-liquid interface.

Epidermis generated in vitro: practical considerations and applications.

The technology for culture of epidermis is one of the most advanced to date for generation of a tissue in vitro. Cultured epidermis is already used for a number of applications ranging from use as a permanent skin replacement to use as an organotypic culture model for toxicity testing and basic research. While simple epidermal sheets have been grafted successfully, more advanced models for skin replacement consisting of both dermal and epidermal components are in development and being tested in a number of laboratories.

The living skin equivalent: Its manufacture, its organotypic properties and its responses to irritants.

A living skin equivalent that resembles human skin has been manufactured as a quality controlled product and commercialized. The skin equivalent consists of a dermal equivalent, reconstituted with collagen and dermal fibroblasts that are biosynthetically active, and a differentiated epidermis that arises from cultured keratinocytes plated onto the surface of the dermal equivalent. The organotypic system is configured so that the dermis is in contact with a nutrient pool while the stratum corneum of the epidermis is exposed to the atmosphere and is thereby accessible for the application of test substances in liquid, gel, emulsion, salve, powder or other forms.