Provided are live, artificial, skin substitute products and methods of making and using the same, such as for wound treatment and compound testing, including compound testing for efficacy, toxicity, penetration, irritation and/or metabolism testing of drug candidates or compositions such as cosmetics. Described herein is an artificial mammalian skin substitute product, comprising: (a) optionally, but in some embodiments preferably, a first (hypodermis-like) layer comprising live mammalian adipocytes (e.g., induced pre-adipocytes) in a first hydrogel carrier; (b) a second (dermis-like) layer contacting or directly contacting the first layer and comprising live mammalian fibroblast cells and' live mammalian follicle dermal papilla cells in combination in a second hydrogel carrier; (c) a third (epidermis-like) layer contacting or directly contacting the second layer (i.e., on the opposite side thereof as the first layer, so that the second layer is sandwiched between the first and third layers when the first layer is present), the third layer comprising live mammalian keratinocytes and live mammalian melanocytes in combination in a third hydrogel carrier.

A method for producing a skin equivalent comprising the steps of: a) producing a dermal matrix containing a first layer and a second layer comprises the steps of i) providing the first layer; ii) providing a polymerizable solution comprising a liquid and at least one polymer; iii) adding the polymerizable solution to the first layer; iv) polymerizing the solution to form the second layer; v) compressing the second layer, wherein as a result of the compression the liquid content of the second layer is reduced and the first layer and second layer are joined together to form the dermal matrix; and vi) introducing at least one cavity into the dermal matrix; b) introducing at least one hair follicle and/or hair follicle germ into the at least one cavity of the dermal matrix produced in step a) to produce a skin surrogate; and c) adding at least one further cell population.

The invention relates to the use of germ cells for obtaining a micro hair follicle and to the use thereof for evaluating the effect of cosmetic, pharmaceutical or dermatological products and also for the prophylactic or therapeutic treatment of a state of reduced pilosity.

The invention relates to a method for manufacturing a bio-ink by additive deposition, which comprises supplying: a first solution including between 5 and 40 wt. % gelatin; a second solution including between 15 and 35 .wt. % alginate; a third solution including between 1 and 15 wt. % fibrinogen, and optionally living cells in suspension; and creating a mixture including: around 35 to 65 vol. % of the first solution; around 15 to 35 vol. % of the second solution; and around 15 to 35 vol. % of the third solution, said proportions being selected so that they add up to 100%. Said bio-ink allows the additive deposition of objects that can be polymerised by means of a solution including calcium ions and thrombin. Said objects can be incubated and can be used as a substitute for body tissue, for example (with added fibroblasts) as skin substitute.

Provided herein are methods for directing differentiation of human pluripotent stem cells into a three-dimensional multilayered skin composition comprising an epidermal layer, a dermal layer, and a plurality of cells capable of forming a functional hair follicle. Also provided herein are three-dimensional, multilayered engineered skin compositions and methods of using the same for drug screening, for screening compounds for effects on hair growth, and for other applications.

The invention relates to the use of germ cells for obtaining a micro hair follicle and to the use thereof for evaluating the effect of cosmetic, pharmaceutical or dermatological products and also for the prophylactic or therapeutic treatment of a state of reduced pilosity.

Epidermis equivalents capable of pigmentation include cells derived from the differentiation of matrix cells; reconstructed skins comprised thereof, optionally containing hair follicles, are useful for evaluating the effect of topical cosmetic, pharmaceutical or dermatological products and may also be used for the preparation of grafts suited to be transplanted on mammals, more particularly on human patients such as victims of third-degree burns.

A method for preparing a bioactive composition containing conditioned cell culture medium is disclosed. The method comprises culturing cells of two or more eukaryotic cell line to form conditioned culture media, separating the cultured cells from the conditioned culture media, and combining conditioned culture media to form a bioactive composition. Novel bioactive compositions, formulations and their use in treating of a variety of diseases and health conditions are also disclosed.

The invention provides a micro-organ composite which comprises a core group of cells and an outer layer of cells, wherein the cells of the core group are mesenchymal cells and the cells of the outer layer are epithelial cells or wherein the cells of the core group are epithelial cells and the cells of the outer layer are mesenchymal cells, and wherein the core group of cells is at least partially encapsulated by the outer layer of cells.

A method for preparing a bioactive composition containing conditioned cell culture medium is disclosed. The method comprises culturing cells of two or more eukaryotic cell line to form conditioned culture media, separating the cultured cells from the conditioned culture media, and combining conditioned culture media to form a bioactive composition. Novel bioactive compositions, formulations and their use in treating of a variety of diseases and health conditions are also disclosed.