Inventors: Rebecca Morris
Publication date: 2008-02-01
Invention summary:
A chemically defined animal cell culture medium composition comprising (a) a synthetic basal medium; (b) calcium at a concentration of from about 1.2 mM to about 1.4 mM; (c) a ratio of sodium to potassium in the range of from about 57.5 to about 27.9; (d) retinoid at a concentration of from about 0.01 mg/L to about 1.0 mg/L; (e) vitamin D at a concentration of from about 0.01 mg/L to about 1.2 mg/L; and (f) linoleic acid or an ester thereof at a concentration of from about 0.01 mg/L to about 1 mg/L
Abstract:
The invention is directed to a chemically defined animal cell culture media, and methods for preparing such a medium, wherein the media are suitable for culturing epidermal cells, preferably human epidermal cells, including cells of the hair follicle. The invention further provides for methods of culturing epidermal cells, hair follicles, and skin explants in the media as well as uses of the cell cultures and explant cultures in screening assays.
First Claim:
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Background:
The epidermis consists of multiple layers of epithelial cells, including keratinocytes. Epidermal keratinocytes can be terminally differentiated, characterized by stratification and desmosome formation, or they can be in a state of growth and proliferation. Keratinocyte stem cells represent a population of cells which can be mobilized to reepithelialize the epidermis or to regenerate the hair follicle. Other follicular cell types include sheath cells, which form the follicular connective tissue, and the follicular papillae cells, which are mesenchymal cells that regulate hair follicle differentiation.
In vitro cultivation of epidermal cells, including hair follicle cells, has been difficult to achieve in the absence of unpurified biological components (such as serum or pituitary extract) or feeder cells to provide an adequate nutritional environment. Chemically defined media that permit in vitro proliferation, expansion and differentiation of murine and human epidermal cells are advantageous for developing physiologically accurate in vitro models of skin and hair growth, thus avoiding the need for animal-based research. Also, chemically defined media are preferred when growing epidermal cells for human therapeutic purposes, such as skin grafts. The use of chemically defined media reduces the risk of adverse effects presently caused by the undefined constituents in serum-supplemented cell culture media and feeder cell layers.
It is important to establish culture conditions which support the growth and differentiation of epidermal and follicular cells, particularly human cells and skin explants used to develop skin grafts. Presently, skin grafts do not contain hair follicles, sebaceous glands or eccrine glands. It is important to develop graft growth conditions which support the development of these epidermal substructures. Hair provides protection for the skin graft and is also important for aesthetic purposes. The incorporation of sebaceous glands into a skin graft will prevent the skin from being dry and flaky. Eccrine (sweat) glands are crucial for proper regulation of the body's response to changes in temperature. |
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