Gas-liquid level growth medium for melanin and keratinocytes for constructing reconstituted skin model

Abstract

The invention provides a gas-liquid level growth medium for melanin and keratinocytes for constructing a reconstituted skin model and relates to the technical field of tissue engineering. Gathered and abnormally germinated melanin model tissue sections prove that the main reason lies in non-uniform melanin germination and excessive germination of the keratinocytes. The growth medium has the characteristics of promoting uniform melanocytes germination and inhibiting excessive germination of the keratinocytes, and attachment and proliferation of melanocytes are promoted by adding cholera toxin and TPA (12-O-tetradecanoylphorbol-13-acetate). In terms of 500 ml, the gas-liquid level growth medium for the melanin and the keratinocytes for constructing the reconstituted skin model comprises 500 ml of cutin serum-free medium K-SFM, 2-30 mu g of insulin, 10-800 mu g of hydrocortisone, 10-200mg of bovine pituitary extract, 20-2000 mu g of an upper epidermal growth factor, 1-1500 mu g of a basic fibroblast growth factor, 5-80 mu g of a keratinocyte growth factor, 20-200 ng of TPA, 10-200 mg of CaCl2 and 20-200 nM of cholera toxin.

Description

technical field [0001] The invention relates to the technical field of tissue engineering, in particular to a melanin and keratinocyte air-liquid surface medium for constructing a recombinant skin model. Background technique [0002] The formation of human skin color is mainly due to the melanosomes produced and secreted by melanocytes reaching keratinocytes and redistributed in keratinocytes. Establishing a co-culture model of human epidermal melanocytes and keratinocytes can provide a research platform for studying the mechanism of skin pigmentation. Under physiological conditions, melanocytes can contact and communicate with surrounding keratinocytes through dendrites, and keratinocytes control the growth, morphology, dendrite formation and antigen expression of melanocytes through direct intercellular connections. [0003] Co-culture of melanocytes and keratinocytes in direct contact can be divided into co-culture of monolayer cells and co-culture of reconstructing thre...

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Problems solved by technology

[0004] At present, the problem of melanin-containing recombinant artificial skin is that the melanin growth is uneven, which leads to the accumulation of dot-shaped melanin or half-moon-shaped melanin on the surface of the melanin model, which is inconsistent with the state of normal human skin, forming a patchy skin, which is very important for melanin. Whitening Efficacy Evaluation of Vegetarian Model Formation Resistance

[0005] CN201019018002.2 The melanocyte and keratinocyte co-cultivation medium used in the pigment-containing artificial skin detection model and its preparation method is added with reducing vitamin C, which has an inhibitory effect on the growth of melanin and is not conducive to Efficacy evaluation of whitening in the later stage, and the survival time of the cultured pigmented artificial skin model is relatively short, only 6 to 11 days, which is not conducive to the evaluation of the efficacy of long-acting whitening products

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