Provided are a composition for preventing hair loss and stimulating hair growth, which includes small-sized stem cells, particularly, small-sized stem cells having a diameter of 8 μm or less, or a culture thereof as an active ingredient, a method of preparing the same, and a use of the same. Provided is a use of the composition utilizing a hair growth stimulating function of the small-sized stem cells having a diameter of 8 μm or less to considerably increase activity of hair follicle stem cells in a telogen phase.

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 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.

The present invention relates to a method for producing hair microfollicles comprising the steps of: a) providing de novo papillae, b) providing other cell populations selected from the group of fibroblasts, keratinocytes and melanocytes, and co-culturing the de novo papillae with at least one other cell population in non-adherent culture vessels. The present invention relates also to methods of producing de novo papillae usable in said method for producing hair microfollicles.

Provided is a hair follicle germ capable of forming a hair shaft-like structure in vitro simply and within a short period of time. A method of producing a hair follicle germ includes: inoculating epithelial cells and mesenchymal cells; maintaining the epithelial cells and the mesenchymal cells in a culture solution in which (a) laminin and entactin, and/or (b) type IV collagen is dispersed; and co-culturing the epithelial cells and the mesenchymal cells in a culture solution to form a hair follicle germ.

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.

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.

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.

Provided are skin constructs and methods of making and using the same, such as for wound treatment and/or compound testing, including compound testing for efficacy, toxicity, immune response, penetration, irritation and/or metabolism testing of drug candidates or compositions such as cosmetics.

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.