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Epidermis skin model with melanin and construction method and application of epidermis skin model

A technology of skin model and melanin, applied in the direction of epidermal cells/skin cells, artificial cell constructs, biochemical equipment and methods, etc., can solve problems such as unfavorable fibroblast stretching and matrix secretion, epidermal layer death, and dermal layer death , achieve the effect of shortening construction time, reducing production cost and simplifying operation process

Pending Publication Date: 2020-05-08
GUANGDONG BOXI BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The skin model constructed by this method contains dermis and epidermis, but the dermis only contains fibroblasts, which is not conducive to the stretching of fibroblasts and matrix secretion, and eventually leads to the death of the dermis, while the epidermis is not directly in contact with the medium , there is a gap between it and the medium, which eventually leads to the death of the epidermis

Method used

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  • Epidermis skin model with melanin and construction method and application of epidermis skin model
  • Epidermis skin model with melanin and construction method and application of epidermis skin model
  • Epidermis skin model with melanin and construction method and application of epidermis skin model

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Embodiment 1

[0055] This embodiment introduces a kind of epidermal skin model containing melanin, the structure of the epidermal skin model containing melanin is stratum corneum, granular layer, spinous cell layer and basal layer successively from top to bottom, and the epidermal skin model containing melanin The melanin-containing epidermal skin model is formed by using a mixture of primary human epidermal keratinocytes and melanocytes through in vitro co-culture, submerged culture and air-liquid surface culture. The epidermal skin model containing melanin provided in this example is constructed by expanding a small amount of seed cells in vitro by applying the principles of cell biology and tissue engineering. Its structure is highly similar to that of normal epidermis, with a top-down structure The stratum corneum, granular layer, spinous layer, and basal layer are in sequence, with a complete epithelial structure and containing melanocytes, which can highly simulate the synthesis, secre...

Embodiment 2

[0059] This embodiment focuses on an in vitro construction method of a melanin-containing epidermal skin model, which includes the following steps:

[0060] Step 1. Obtain primary human epidermal keratinocytes and melanocytes:

[0061] Soak the isolated human skin tissue in ethanol with a mass fraction of 75% for 30 seconds, rinse it with phosphate buffered saline (PBS), spread the epidermis of the tissue on a plate, peel off the subcutaneous loose connective tissue with elbow scissors, and expose the dense Connective tissue, then transferred to fresh PBS to soak and clean, and then immersed the tissue in 1.2U / mL Dispase enzyme digestion solution with a mass fraction of 1%, soaked at 37°C for 2 hours, and digested in 37°C trypsin / EDTA Digest for 10 minutes at 37°C, add 15 mL of stop solution, gently pipette 100 times with an elbow pipette, filter with a 200-mesh sieve under sterile conditions, centrifuge at 800 rpm for 6 minutes, discard the supernatant, and obtain the primary...

Embodiment 3

[0074] This embodiment focuses on an in vitro construction method of a melanin-containing epidermal skin model, which includes the following steps:

[0075] Step 1. Obtain primary human epidermal keratinocytes and melanocytes:

[0076] Soak the isolated human skin tissue in ethanol with a mass fraction of 75% for 30 seconds, rinse it with phosphate buffered saline (PBS), spread the epidermis of the tissue on a plate, peel off the subcutaneous loose connective tissue with elbow scissors, and expose the dense Connective tissue, then transferred to fresh PBS to soak and clean, and then immersed the tissue in 1.2U / mL Dispase enzyme digestion solution with a mass fraction of 1%, soaked at 37°C for 2 hours, and digested in 37°C trypsin / EDTA Digest for 12 minutes at 37°C, add 15 mL of stop solution, gently pipette 150 times with an elbow pipette, filter with a 200-mesh sieve under sterile conditions, centrifuge at 800 rpm for 6 minutes, discard the supernatant, and obtain the primary...

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Abstract

The invention provides an epidermis skin model with melanin. The epidermis skin model with melanin structurally consists of stratum corneum, a granular layer, a prickle cell layer and a basal layer from top to bottom. The epidermis skin model with melanin contains melanin cells, and the epidermis skin model with melanin is obtained from a mixture of original human epiderm keratinocytes and melanincells through in-vitro coculture, submerged culture and gas-liquid level culture.

Description

technical field [0001] The invention belongs to the field of tissue engineering biomaterials, in particular to a melanin-containing epidermal skin model and its construction method and application. Background technique [0002] In biomedical research, cosmetics, pharmaceuticals, chemicals and other products must undergo a large number of tests before they go on the market to ensure the safety and efficacy of the products. Animal experiments have always been the main mode in this research field, but due to the species differences between animals and humans, experimental animals cannot accurately simulate human skin, which leads to deviations in experimental results. And because of the 3R principle (reduce, optimize and replace), researchers are prompted to find a more suitable detection method. At present, in vitro alternative methods have become the main mode of modern toxicology research system and one of the new directions of biomedical development, accepted and implement...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0629C12N5/0626C12N5/0698C12N2500/90C12N2501/585C12N2500/30C12N2501/30C12N2500/05Y02A50/30
Inventor 杨文娟张晓娥卢永波张勇杰
Owner GUANGDONG BOXI BIO TECH CO LTD
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