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 even

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

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0054] Example 1:

[0055] This embodiment introduces a melanin-containing epidermal skin model. The melanin-containing epidermal skin model has a stratum corneum, a granular layer, a spinous cell layer, and a basal layer from top to bottom. The melanin-containing epidermal skin model It contains melanocytes, and the melanin-containing epidermal skin model adopts a mixture of primary human epidermal keratinocytes and melanocytes through in vitro co-culture, sub-liquid culture and air-liquid surface culture. The melanin-containing epidermal skin model provided in this embodiment is constructed by applying the principles of cell biology and tissue engineering to a small number of seed cells after being expanded in vitro. Its structure is highly similar to the normal epidermal structure, with a top-down structure The stratum corneum, the granular layer, the spinous layer and the basal layer are in sequence. They have a complete epithelial structure and contain melanocytes, which can...

Example Embodiment

[0058] Example 2

[0059] This embodiment focuses on an in vitro construction method of an epidermal skin model containing melanin, 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 with phosphate buffered saline PBS, spread the tissue epidermis face down in a petri dish, peel off the subcutaneous loose connective tissue with elbow scissors, and expose it to dense The connective tissues are then transferred to fresh PBS to soak and clean, then the tissues are immersed in 1.2U / mL Dispase enzyme digestion solution with a mass fraction of 1%, soaked at 37°C for 2 hours, and put into 37°C trypsin / EDTA digestion Digest the solution at 37°C for 10 minutes, add 15 mL of stop solution, gently pipette 100 times with an elbow pipette, filter with a 200-mesh sieve under aseptic conditions, centrifuge at 800 rpm for 6 minutes,...

Example Embodiment

[0073] Example 3:

[0074] This embodiment focuses on an in vitro construction method of an epidermal skin model containing melanin, 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 with phosphate buffered saline PBS, spread the tissue epidermis face down in a petri dish, peel off the subcutaneous loose connective tissue with elbow scissors, and expose it to dense The connective tissue is then transferred to fresh PBS and soaked and cleaned. Then the tissue is immersed in 1.2U / mL Dispase digestion solution with a mass fraction of 1%, soaked at 37°C for 2 hours, and put into 37°C trypsin / EDTA digestion Digest the solution at 37°C for 12 minutes, add 15 mL stop solution, gently pipette 150 times with an elbow pipette, filter with a 200-mesh sieve under aseptic conditions, centrifuge at 800 rpm for 6 minutes, discard...

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