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Simultaneous autologous epidermic cell and fibrocyte preparation method and biological cosmetic product thereof

A technology of fibroblasts and epidermal cells, applied in the directions of medical preparations, animal cells, cosmetic preparations containing active ingredients, etc., can solve the problems of poor effect, insufficient skin beauty effect, increased usage, etc., and achieve remarkable results. , restore elasticity and luster, shrink pores

Inactive Publication Date: 2015-12-23
怡诺博(北京)生物医学技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The application of these two factors tends to produce obvious dependence. As the number of injections increases, people's sensitivity to them decreases, which leads to patients using this method more frequently, and the amount of each use will gradually increase.
[0005] To sum up, due to various reasons and problems, biological beauty products currently on the market have various ineffective effects, and the curative effect on people’s skin beauty is far from enough, even if various nutrients are added to them

Method used

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  • Simultaneous autologous epidermic cell and fibrocyte preparation method and biological cosmetic product thereof
  • Simultaneous autologous epidermic cell and fibrocyte preparation method and biological cosmetic product thereof
  • Simultaneous autologous epidermic cell and fibrocyte preparation method and biological cosmetic product thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1 Preparation of human autologous epidermal cells

[0055] By taking people 0.1-2cm 2 For the skin, infiltrate 5ml of 1×phosphate buffered saline (PBS, pH 7.4, containing 100IU / ml penicillin and 0.1mg / ml streptomycin) for 15 minutes, suck it up with a 1ml pipette gun, and then rinse with 1×PBS (pH Value 7.4, containing 100IU / ml penicillin and 0.1mg / ml streptomycin) were also washed repeatedly, digested with disperse II after washing, placed in a refrigerator at 4°C overnight, and the skin was taken out the next day and the epidermis and dermis were torn apart with tweezers.

[0056] Cut the epidermis to 1mm with ophthalmic scissors 3 After washing with 1×PBS, add it to a 15ml centrifuge tube, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, add 3ml of 0.25% (mass volume ratio) trypsin to digest for 30 minutes, wherein the trypsin contains mass volume Ratio of 0.01% EDTA; pass through a 40μm cell sieve after digestion, add 10ml 1×PBS after filt...

Embodiment 2

[0057] Embodiment 2 Preparation of human autologous fibroblasts

[0058] Get the dermis obtained in Preparation Example 1 and cut it to 1mm with ophthalmic scissors 3 After washing with 1×PBS, add it to a 15ml centrifuge tube, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, add 3ml of 0.25% trypsin to digest for 30 minutes, pass through a 40μm cell sieve after digestion, add 10ml1 after filtration ×PBS, centrifuged at 1000 rpm for 10 minutes to obtain a single cell pellet, resuspended in 10ml of 1×PBS again, centrifuged at 1000 rpm for 10 minutes to obtain a single cell pellet, added α-MEM medium to resuspend, and counted , with α-MEM medium (containing 10% fetal bovine serum, 50ng / ml recombinant human stem cell growth factor, 25ng / ml recombinant human epidermal growth factor and 25ng / ml recombinant human basic fibroblast growth factor) according to 1-10 ×10 6 Cells / ml add 15ml to 75cm 2 culture flask at 37°C, 5% CO 2 Cultivate in an incubator; replace with ...

Embodiment 3

[0060] Example 3 Human autologous epidermal cells and fibroblast-specific protein expression detection

[0061]Take 1 part of 50 μL each of the autologous epidermal cells prepared in Example 1 and the fibroblast suspension prepared in Example 2 (2×10 6 1) Add 500 μL of pre-cooled ready-to-use total protein extraction and lysate (RIPA, Beijing Biotech Company) (containing 1% protease inhibitor by volume, Roche Company, Switzerland), incubate on ice for 20 minutes, 13000 rpm, 4 Centrifuge for 20 minutes. Take the supernatant, aliquot and store at -80°C for later use. The protein concentration was determined according to the instructions of the bicinchoninic acid (BCA) protein quantification kit (Wuhan Boster Company). Adjust the protein concentration with RIPA, the final concentration of the sample is 1.0 μg / μL, add 5× protein sample buffer, and keep at 95°C for 5 minutes.

[0062] According to the molecular weight of the target protein, a 10% separating gel was prepared, and...

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Abstract

The invention provides a simultaneous autologous epidermic cell and fibrocyte preparation method. The simultaneous autologous epidermic cell and fibroblast preparation method comprises the steps of obtaining epidermic cells from the autologous skin and performing cultivation to obtain fibrocyte. The invention further provides a biological cosmetic product containing the autologous epidermic cells and / or the fibrocyte. The biological cosmetic product containing the autologous living cells is produced by adding one or two types of cells into plasma rich of platelets in autologous peripheral blood and meanwhile adding nutritional factors. The biological cosmetic product can have the cosmetic effects of removing freckles caused by leucoderma, healing scars, removing skin wrinkles, removing freckles and the like.

Description

technical field [0001] The invention relates to a method for simultaneous preparation of autologous epidermal cells and fibroblasts and a biocosmetic product obtained by the method. In particular, the present invention relates to fibroblasts obtained by obtaining epidermal cells from autologous skin and culturing them, adding one or two types of cells to the platelet-rich plasma of autologous peripheral blood, and at the same time adding nutritional factors to produce The bio-cosmetic product containing autologous living cells can be used for cosmetic effects such as vitiligo freckle removal, scar repair, skin wrinkle removal and freckle removal. Background technique [0002] A few days ago, a variety of cells used in beauty products have appeared on the market, and have been endowed with various magical effects and hopes by merchants, especially the application of stem cells in beauty. In fact, living cells have achieved some good results in the application of medical cosm...

Claims

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Application Information

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IPC IPC(8): C12N5/071A61K8/98A61K35/33A61K35/12A61Q19/00A61Q19/02A61Q19/08A61P17/02
Inventor 黄启明王旭王玉东
Owner 怡诺博(北京)生物医学技术有限公司
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