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Method for constructing tissue engineering epidermis for detecting irritation of cosmetics on polycarbonate film

A technology of polycarbonate membrane and tissue engineering, applied in measuring devices, medical science, instruments, etc., can solve the problems of high price and limited sources, and achieve the effects of simple operation, convenient synthesis and large-scale production, and low cost

Inactive Publication Date: 2014-06-25
PROYA COSMETICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The European Center for the Validation of Alternative Methods (ECVAM) issued the "Technical Standards for Human Skin Models for In Vitro Skin Irritation Tests" in May 2007, and the "Two Guidelines" in November 2008. In the statement "Verification Method for In Vitro Skin Irritation Test", it has been indicated that tissue engineered skin can be used as an alternative model for in vitro skin irritation test, such as tissue engineered skin Episkin®, EpiDerm® and SkinEthic®, etc., but these commercial products are expensive , sources are limited and other defects, so it is relatively cheap to construct a tissue-engineered skin model that can be used for skin irritation tests under laboratory conditions, which is convenient for mass production and can effectively solve these problems

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] A method for constructing a tissue-engineered epidermis on a polycarbonate membrane for the detection of cosmetic irritation using the following steps:

[0015] (1) Isolation and culture of epidermis-forming cells: 16cm2 ​​of the back skin of the induced fetus was taken under sterile conditions, rolled into a 0.5cm×0.5cm piece of skin, washed with normal saline, and 30ml with 0.125% mass percentage Dispase II enzyme was digested at 20°C for 10 hours, then 10ml of 0.1% trypsin by mass was added and digested at 37°C for 30 minutes to obtain a solution of epidermal cells separated into single cells, centrifuged at 5000rpm for 10 minutes, the supernatant was removed, and washed with normal saline The lower layer of cells was then centrifuged at 5000rpm for 10 minutes. After repeated washing and centrifugation for 3 times, 50ml of restricted epidermal cell serum-free culture (DK-SFM) containing 30U / ml penicillin and 30ug / ml streptomycin was added. Incubate at ℃ for 24 hours ...

Embodiment 2

[0019] A method for constructing a tissue-engineered epidermis on a polycarbonate membrane for the detection of cosmetic irritation using the following steps:

[0020] (1) Isolation and culture of epidermis-forming cells: 16cm2 ​​of the back skin of the induced fetus was taken under sterile conditions, rolled into a 0.5cm×0.5cm piece of skin, washed with normal saline, and 30ml with 0.125% mass percentage Dispase II enzyme was digested at 20°C for 10 hours, then 10ml of 0.1% trypsin by mass was added and digested at 37°C for 30 minutes to obtain a solution of epidermal cells separated into single cells, centrifuged at 5000rpm for 10 minutes, the supernatant was removed, and washed with normal saline The lower layer of cells was then centrifuged at 5000rpm for 10 minutes. After repeated washing and centrifugation for 3 times, 50ml of restricted epidermal cell serum-free culture (DK-SFM) containing 35U / ml penicillin and 35ug / ml streptomycin was added. Incubate at ℃ for 24 hours ...

Embodiment 3

[0024] A method for constructing a tissue-engineered epidermis on a polycarbonate membrane for the detection of cosmetic irritation using the following steps:

[0025] (1) Isolation and culture of epidermis-forming cells: 16cm2 ​​of the back skin of the induced fetus was taken under sterile conditions, rolled into a 0.5cm×0.5cm piece of skin, washed with normal saline, and 30ml with 0.125% mass percentage Dispase II enzyme was digested at 20°C for 10 hours, then 10ml of 0.1% trypsin by mass was added and digested at 37°C for 30 minutes to obtain a solution of epidermal cells separated into single cells, centrifuged at 5000rpm for 10 minutes, the supernatant was removed, and washed with normal saline The lower layer of cells was then centrifuged at 5000rpm for 10 minutes. After repeated washing and centrifugation for 3 times, 50ml of restricted epidermal cell serum-free culture (DK-SFM) containing 40U / ml penicillin and 40ug / ml streptomycin was added. Cultivate for 24 hours at ℃...

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Abstract

The invention relates to a method for constructing a tissue engineering epidermis for detecting the irritation of cosmetics on a polycarbonate film. The method is characterized by comprising the following steps: (1) carrying out separated culture on epidermization cells; (2) preparing a lipid mixture solution; and (3) constructing the tissue engineering epidermis on the polycarbonate film. The method has the advantages that the operation is easy and convenient and the cost is low; the method is good for synthesis and large-scale production of a product and the product has a basal layer, a spinous layer, a granular layer, a transparent layer and the cuticle tissue engineering epidermises which are in completely differentiated shapes, and the irritation of the cosmetics can be conveniently detected.

Description

technical field [0001] The present invention relates to a method for constructing a tissue-engineered epidermis on a polycarbonate membrane for detection of cosmetic irritation. Background technique [0002] Traditional skin irritation and corrosion tests are mostly carried out with animals. A single in vivo skin irritation test is generally carried out with rabbits, and at least 4 healthy adult animals are selected for each test. If an irritant response is expected, one animal should be considered for the initial test. If there is no obvious positive reaction (erythema or edema score greater than 2), at least 2 more animals should be used for the test. If there is no response as expected, 3 animals can be used for the initial test. After using at least 3 animals, if the reaction is still suspected or unclear, a retest should be considered. Multiple in vivo skin irritation test refers to the repeated skin exposure of experimental animals within 7-14 days, which is more e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531A61L27/60A61L27/38A61L27/18A61L27/24A61L27/54
CPCG01N33/5008
Inventor 毕永贤杨盼盼陈斌
Owner PROYA COSMETICS
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