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

A skin model and culture medium technology, applied in cell culture active agents, epidermal cells/skin cells, tissue culture, etc., can solve the problems of inconsistent state, unfavorable whitening effect evaluation, short survival time of artificial skin model, etc., to improve accuracy The effect of degree and precision

Inactive Publication Date: 2017-11-07
GUANGDONG BOXI BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 keratinocyt

Method used

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

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Embodiment 1

[0021] Acquisition of cells used in the construction Acquisition of cells used in the construction

[0022] 1. Acquisition and subculture of primary melanocytes: Melanocytes can be derived from circumcised normal skin tissue. For details on the acquisition and subculture, please refer to "Research on the Construction of Tissue Engineering Skin Containing Melanocytes" by Liu Yuan;

[0023] 2. Acquisition and subculture of keratinocytes: Keratinocytes can be derived from normal skin tissue after circumcision. For details on the acquisition and subculture, please refer to "Isolation, Purification, Culture and Identification of Human Epidermal Stem Cells" by Ma Yingzhi;

[0024] Keratinocyte serum-free medium K-SFM is used as the submerged medium for the co-culture of keratinocytes and melanocytes;

[0025] Preparation of air-liquid surface medium for co-culture of keratinocytes and melanocytes

[0026] a. The preparation of the medium on the first day of the gas-liquid level: us...

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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...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0626C12N5/0629C12N2500/14C12N2500/36C12N2500/84C12N2501/11C12N2501/115C12N2501/117C12N2501/33
Inventor 李芳卢永波
Owner GUANGDONG BOXI BIO TECH CO LTD
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