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In vitro recombinant human skin epidermis model and its preparation method and application

A human skin and external recombination technology, applied in biochemical equipment and methods, epidermal cells/skin cells, tissue screening, etc., can solve the problems of easy shrinkage of the model, defects in the link structure, and model shedding, etc., to increase the surface area and improve The effect of filter rate

Active Publication Date: 2020-05-19
GUANGDONG BOXI BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During the in vitro construction process, the model is prone to shrinkage and other phenomena, and the link structure between the epidermal model and the dermal matrix is ​​defective, which may easily lead to the model falling off, etc.
Defects in these aspects lead to the imperfect barrier function of the existing recombinant epidermal model, high permeability to the test substance or leakage, which affects the accuracy of the in vitro safety test results. For example, Epiderm and Episkin are Two animal surrogate models for in vitro skin irritation prediction approved by the OECD, but there are certain misjudgments in the judgment of the irritation standard

Method used

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  • In vitro recombinant human skin epidermis model and its preparation method and application
  • In vitro recombinant human skin epidermis model and its preparation method and application
  • In vitro recombinant human skin epidermis model and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] The in vitro recombinant human skin epidermis model in this implementation is composed of basal cell layer, spinous cell layer, granule cell layer and keratinocyte layer. The lower surface of the basal cell layer has a layer of hemidesmosomal protein, and the upper surface of the basal cell layer is covered with a spinous cell layer, which is composed of 7 layers of spiny cells with short protrusions, and the upper surface of the spinous cell layer The surface is covered with a layer of granule cells, which consists of three layers of cells containing transparent granules, and the upper surface of the layer of granule cells is covered with a layer of corneocytes. The thickness of the in vitro recombinant human skin epidermis model in this embodiment is 120 μm.

[0103] The preparation method of the above-mentioned in vitro recombinant human skin epidermis model consists of the following steps:

[0104] 1. Preparation of Matrigel Microparticles

[0105] Dilute Matrigel...

Embodiment 2

[0124] The in vitro recombinant human skin epidermis model in this implementation is composed of basal cell layer, spinous cell layer, granule cell layer and keratinocyte layer. The lower surface of the basal cell layer has a layer of hemidesmosomal protein, and the upper surface of the basal cell layer is covered with a spinous cell layer, which is composed of 6 layers of spiny cells with short protrusions, and the upper surface of the spinous cell layer The surface is covered with a layer of granule cells, which consists of three layers of cells containing transparent granules, and the upper surface of the layer of granule cells is covered with a layer of corneocytes. The thickness of the in vitro recombinant human skin epidermis model in this embodiment is 120 μm.

[0125] Its preparation method is as follows:

[0126] 1. Preparation of Matrigel Microparticles

[0127] This step is the same as in Example 1.

[0128] 2. Screen and expand epidermal stem cells

[0129] Imm...

Embodiment 3

[0142] The in vitro recombinant human skin epidermis model in this implementation is composed of basal cell layer, spinous cell layer, granule cell layer and keratinocyte layer. The lower surface of the basal cell layer has a layer of hemidesmosomal protein, and the upper surface of the basal cell layer is covered with a spinous cell layer, which is composed of 8 layers of spiny cells with short protrusions, and the upper surface of the spinous cell layer The surface is covered with a granule cell layer, which is composed of 4 layers of cells containing transparent granules, and the upper surface of the granule cell layer is covered with a keratinocyte layer. The thickness of the in vitro recombinant human skin epidermis model in this embodiment is 150 μm.

[0143] Its preparation method is as follows:

[0144] 1. Preparation of Matrigel Microparticles

[0145] This step is the same as in Example 1.

[0146] 2. Screen and expand epidermal stem cells

[0147] Immerse the is...

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Abstract

The invention discloses an in-vitro recombined human skin epidermis model.A layer of hemidesmosome protein is arranged on the lower surface of a basal cell layer, the upper surface of the basal cell layer is covered with 6-8 spinous cell layers containing short protruding spinous cells, the upper surface of the spinous cell layers is covered with 3-4 granular cell layers containing transparent granular cells, the upper surface of the granular cell layers is covered with a keratinocyte layer, all the cell layers are obviously separated, and the overall barrier function of a lipid composition is approximate to that of human skin.A preparation method comprises the steps of preparation of matrigel particles, screening and amplification of epidermal stem cells, preparation of the basal cell layer, preparation of the spinous cell layers, preparation of the granular cell layers, and maturation and keratinization of the in-vitro recombined epidermis model.The model is applied to evaluation of cosmetics with the human skin barrier repair effect, screening of plant extract with the human skin barrier repair effect, evaluation and detection of irritation of medical apparatus materials on the human skin and detection of irritation of chemicals on the skin.

Description

technical field [0001] The invention belongs to the technical field of biological materials, and in particular relates to a human epidermis model. Background technique [0002] In the field of biomedical research, animal experiments have been used as the main research and verification model for the safety evaluation of skin care products, contact materials, and skin care products. Due to inter-species differences, experimental animals cannot accurately replicate the complex biological and physiological characteristics exhibited by human skin, resulting in biased and one-sided experimental results, and even erroneous conclusions. With the implementation of EU animal welfare and 3R (reduce, optimize and replace) principles, in vitro alternative methods have become the main mode of modern toxicology research system and one of the new directions of biomedical development, accepted by European governments and international regulations and implement. Animal experiments have even...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12Q1/02
CPCC12N5/0698C12N2502/09C12N2503/06G01N33/5008
Inventor 李潇卢永波
Owner GUANGDONG BOXI BIO TECH CO LTD
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