Methods for the efficient isolation and use of pluripotent adipose-derived stem cells (PASCs) are provided. In certain embodiments the methods involve providing an adipose tissue sample from which the stromal vascular fraction is co-cultured with the adipocyte fraction. PASCs can be isolated with a high degree of purification without requiring an additional cell enrichment process (e.g. cell sorting). PASCs and their conditioned media can be used for tissue regeneration within hours of harvesting the adipose tissue, and without requiring cell expansion. PASCs can grow as floating individual cells, as clusters of cells, or attached to surface(s) of the culture vessel. PASCs do not produce teratomas in vivo, nor do they induce immunorejection upon transplantation, and they achieve a high efficiency in grafting. The cells and compositions can be used for cell therapy and to screen new drugs.

Provided are a pharmaceutical composition for regenerating skin, a pharmaceutical composition for improving wrinkles, a pharmaceutical composition for treating wounds, a quasi-drug composition for regenerating skin, a quasi-drug composition for improving wrinkles, a quasi-drug composition for treating wounds, a cosmetic composition for regenerating skin, a cosmetic composition for improving wrinkles, a cosmetic composition for improving wounds, and a medium composition for culturing fibroblasts, each composition including GDF11 or a human-derived adult stem cell culture medium including the same, a method of culturing fibroblasts by using the medium composition, and a method of preparing GDF11 by culturing stem cells. The GDF11 provided in the present invention may be included in a human-derived adult stem cell culture medium to exhibit an effect of promoting fibroblast proliferation, and may thereby be widely applied to the development of a variety of products for skin regeneration, wrinkle improvement, or wound treatment.

An exosome production promoter that promotes production of exosomes from adipose-derived mesenchymal stem cells is provided. The exosome production promoter promotes production of exosomes from adipose-derived mesenchymal stem cells cultured in a serum-free medium and contains EGF. It is preferable that the exosome production promoter further contains IL-1? and trehalose. It is desirable that the exosome production promoter does not contain SCGF. It is preferable that the adipose-derived mesenchymal stem cells are cultured using a nonwoven fabric sheet as a scaffold. The exosomes mass-produced by the technique according to the present invention can be used, for example, for cosmetic plastic procedures or the like.

Methods for the efficient isolation and use of pluripotent adipose-derived stem cells (PASCs) are provided. In certain embodiments the methods involve providing an adipose tissue sample from which the stromal vascular fraction is co-cultured with the adipocyte fraction. PASCs can be isolated with a high degree of purification without requiring an additional cell enrichment process (e.g. cell sorting). PASCs and their conditioned media can be used for tissue regeneration within hours of harvesting the adipose tissue, and without requiring cell expansion. PASCs can grow as floating individual cells, as clusters of cells, or attached to surface(s) of the culture vessel. PASCs do not produce teratomas in vivo, nor do they induce immunorejection upon transplantation, and they achieve a high efficiency in grafting. The cells and compositions can be used for cell therapy and to screen new drugs.

Pharmaceutical compositions for promoting hair follicle regeneration and their respective preparation methods are provided to overcome the ineffectiveness and low efficiency of the conventional hair follicle regeneration techniques. The methods for preparing the pharmaceutical compositions for promoting hair follicle regeneration include seeding dermal papilla cells and adipocyte lineage cells on a culture material in order for the dermal papilla cells and the adipocyte lineage cells to jointly assemble cell spheres. The pharmaceutical compositions can effectively enhance the effect and efficiency of hair follicle regeneration and thereby promote the regeneration of hair follicles.

A cosmetic composition for skin whitening, wrinkle improvement or skin regeneration includes, as an active ingredient, exosomes derived from stem cells comprising proliferating stem cells.

Methods of culturing mesenchymal stem cells are provided. The methods comprise culturing MSCs in a medium comprising nicotinamide and fibroblast growth factor 4 (FGF4). Populations of mesenchymal stem cells generated using the methods described herein and uses thereof are also provided.

A method of generating a population of cells useful for treating a nerve disease or disorder in a subject, the method comprising up-regulating a level of at least one exogenous miRNA in mesenchymal stem cells (MSCs) and/or down-regulating a level of at least one miRNA using a polynucleotide agent that hybridizes to the miRNA, thereby generating the population of cells useful for treating the nerve disease or disorder. Isolated populations of cells with an astrocytic phenotype generated thereby and uses thereof are also provided.

The present disclosure discloses a method for facilitating functions and characteristics of corneal endothelial cells, comprising the following steps of: separating and culturing human orbital adipose-derived stem cells, and extracting a conditioned culture medium; separating and culturing primary human corneal endothelial cells; adding the conditioned culture medium in a basal culture medium for the human corneal endothelial cells, and culturing and proliferating the human corneal endothelial cells. In the present disclosure, the human corneal endothelial cells cultured by the conditioned culture medium extracted from human orbital adipose-derived stem cells have high adherence and proliferation capacities. Human corneal endothelial cells cultured in vitro can be sub-cultured over 10 generations. The proliferation multiple is higher and the morphology and functions of the human corneal endothelial cells can be maintained. Experiments on animals have proved that the human corneal endothelial cells cultured in vitro have excellent cell repair effects.

Methods of producing stem cell conditioned media to treat mammalian injuries or insults. In at least one embodiment of a method of producing a stem cell conditioned media of the present disclosure, the method comprises the steps of culturing at least one stem cell in a first cell culture medium, replacing some or all of the first cell culture medium with a second cell culture medium and further culturing the at least one stem cell in the second cell culture medium, and collecting a quantity of the second cell culture medium after a culture duration, wherein the quantity of the second cell culture medium contains a cell culture byproduct effective to treat a mammalian insult or injury.