Method for building full-thickness skin models

A full-thickness skin model and construction method technology, applied to artificial cell constructs, vertebrate cells, animal cells, etc., can solve problems affecting mass transfer efficiency, separation of culture vessel walls, shrinkage of artificial skin tissues, etc., to ensure vitality and biological functions, avoid non-growth and degradation, increase the effect of thickness

Inactive Publication Date: 2015-06-24
GUANGDONG BOXI BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the artificial skin obtained by this method cannot completely replace skin tissue to meet the test and evaluation requirements for cosmetics, small molecular compounds, active proteins, medical devices, chemicals, and skin-contact materials.
The skin model constructed with scaffold materials and living cells is accompanied by the secretion of collagen of the living cells and the degradation of the scaffold materials during the culture process, resulting in shrinkage of the artificial skin tissue, which seriously affects the mass transfer efficiency in the

Method used

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  • Method for building full-thickness skin models
  • Method for building full-thickness skin models

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Embodiment 1. A kind of construction method of full-thickness skin model comprises the following steps:

[0039] Step 1. Isolation and culture of fibroblasts and epidermal cells: fibroblasts are separated and cultured, and the foreskin tissue that has been removed from fat and connective tissue is cut into 1mm 3 Place the skin tissue containing collagenase 200U·ml -1 , hyaluronic acid 300U·ml -1 In the culture medium I, place it at 4°C overnight, then add an equal volume of digestion solution, stop the digestion after 20 minutes on an air shaker at 37°C at 180 rpm; collect cells by centrifugation at 1000 rpm for 10 minutes; adjust the cell density to 2×10 with culture medium II 5 piece / cm 2 Cell seeding, 5.0% CO 2 , Cultivate at 37°C;

[0040] Epidermal cells were isolated and cultured, and the epidermal tissue was immersed in 1.2U / mL digestion solution, and placed at 4°C for 20 hours; the epidermis was torn off under sterile conditions, and the epidermis was placed...

Embodiment 2

[0049] Embodiment 2. A kind of construction method of full-thickness skin model

[0050] Step 1. Isolation and culture of fibroblasts and epidermal cells

[0051] Isolation and culture of fibroblasts: cut the foreskin piece from which fat and connective tissue has been removed into 1mm 3 Place the skin tissue containing collagenase 200U·ml -1 , hyaluronic acid 300U·ml -1 In culture solution I, put it at 4°C overnight, then add an equal volume of digestion solution, stop digestion after 20 minutes on an air shaker at 37°C at 180 rpm; collect cells by centrifugation at 1000 rpm for 10 minutes; adjust the cell density to 4×10 5 piece / cm 2 Cell seeding, 5% CO 2 , Cultivated under humidity conditions of 37°C.

[0052] Isolation and culture of epidermal cells: immerse the epidermal tissue in 1.2U / mL digestive solution, and place it at 4°C for 20 hours; tear off the epidermis under sterile conditions, and put the epidermal layer into the preheated digestive solution at 37°C , a...

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Abstract

The invention relates to a method for building full-thickness skin models. The full-thickness skin models comprise corium layers and epidermal layers. The corium layers comprise fibroblasts, the epidermal layers comprise epidermal cells, and substances such as cosmetics can be detected on the full-thickness skin models instead of the skin tissues of animals. The method has the advantages that cells of the corium layers are inoculated in gradient time, and can secrete a large quantity of collagen and form mature extracellular matrixes under the joint effects of V<c> (vitamin C), natural three-dimensional supports and growth spaces can be provided for the cells by the extracellular matrixes in cell re-layering procedures, and the biological activity and the biocompatibility of the cells can be improved; repeated inoculation and cell proliferation re-layering procedures are combined with one another, so that the thicknesses of the corium layers can be increased, and the mechanical strength and the stability of the corium layers can be improved; requirements on displaying the activity and biochemical functions of the cells of the corium layers and the epidermal layers can be met by nutrient solution after the corium layers and the epidermal layers are combined with one another.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering medical biomaterials, and specifically relates to a safety and efficacy test for cosmetics, small molecular compounds, active proteins, medical devices, chemicals, disposable hygiene products, etc. A method for constructing a full-thickness skin model for safety and efficacy evaluation of skin-contact materials. Background technique [0002] Skin model refers to the skin substitute artificially prepared in vitro by using the principles and methods of engineering and cell biology. The full-thickness skin model is highly similar to the normal skin structure, with a complete epidermis and dermis structure. In addition to being used to repair and replace defective skin tissue, its greatest use is to replace human skin tissue for cosmetics, small molecular compounds, and active ingredients. Safety and efficacy evaluation of proteins, medical devices, chemicals, disposable hygiene products, ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 明磊国张勇杰卢永波
Owner GUANGDONG BOXI BIO TECH CO LTD
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