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Preparation method for applying autogenous skin fibroblasts to beautifying and anti-aging

A fibroblast and skin technology, applied in the field of biological cells, can solve problems such as allergies, limited effects, and no biologically active substances, and achieve the effect of ensuring the safety of use

Inactive Publication Date: 2016-12-07
ANHUI HUIEN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the existing fine chemical skin care products contain substances that are harmful to the skin such as lead, mercury, alcohol, preservatives, hormones, spices, etc. When using beauty care products, they also cause greater damage to the skin, and even The resulting stains, red blood streaks, allergies and other problems pose a great threat to people's quality of life and health
[0004] Most of the existing beauty care products are made by chemical methods, and there are many harmful substances in the products, and long-term use will have adverse effects on the skin
Moreover, most of the existing products do not have biologically active substances, which cannot effectively promote the metabolism of epidermal cells and improve the luster of the skin.
Moreover, some substances contained in chemical skin care products will cause allergic reactions to the human body, so they are not suitable for all people.
[0005] At present, the bio-cosmetic facial masks on the market have very limited effects on skin beauty, such as moisturizing, whitening and oil control. In order to have better effects on the skin such as wrinkle removal, acne scar and scar repair, spot removal and anti-oxidation, Utilizing the safety and better biological activity of autologous skin fibroblasts for beauty and anti-aging, the market still cannot meet the demand

Method used

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Experimental program
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Effect test

Embodiment 1

[0022] A method for preparing autologous skin fibroblasts for beauty and anti-aging, first surgically cut out 3cm 2 Human abdominal skin, placed in PBS containing 110U / ml ampicillin and 110mg / ml streptomycin sulfate, rinsed with PBS repeatedly for 3 times, then cut off fat and connective tissue;

[0023] Cut the skin to 1.2mm 3 The left and right small pieces were added to DMEM containing 200 U / ml collagenase and 300 U / ml hyaluronidase, and placed overnight (12h) at 4°C;

[0024] Add PBS digestion solution containing 0.25% trypsin and 0.02% EDTA at a ratio of 1:1, centrifuge at 37°C and 180rpm for 20min, and stop the serum; then collect the cells under centrifugation at 1000rpm for 10min; The α-MEM medium containing 15% human autologous serum was used to adjust the cell density to 4×10 5 Cell seeding, 37 °C, 5% CO 2 , open and cultivate for 48 hours under 90% moderate conditions;

[0025] When the cultured skin fibroblasts grow into a monolayer, rinse once with PBS without...

Embodiment 2

[0029] A method for preparing autologous skin fibroblasts for beauty and anti-aging. First, surgically cut 2cm 2 Human abdominal skin, placed in PBS containing 100U / ml ampicillin and 100mg / ml streptomycin sulfate, rinsed with PBS for 5 times, then cut off fat and connective tissue;

[0030] Cut the skin to 0.8mm 3 Add the left and right small pieces into DMEM containing collagenase 200U / ml and hyaluronidase 300U / ml, and place at 4°C overnight; add PBS digestion solution containing 0.25% trypsin and 0.02% EDTA at a ratio of 1:1, 37 ℃, 180rpm air shaker for 20min, the serum was terminated; the cells were collected by centrifugation at 1000rpm for 10min; after DMEM elution twice, the cell density was adjusted with α-MEM culture medium containing 15% human autologous serum at 2×10 per ml 5 Cell seeding, 37 °C, 5% CO 2 , open and cultivate for 48 hours under 90% moderate conditions;

[0031] When the cultured skin fibroblasts grow into a single layer, rinse once with PBS without...

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Abstract

The invention discloses a preparation method for applying autogenous skin fibroblasts to beautifying and anti-aging. The method includes the following steps that the human autogenous abdominal skin is cut and put into PBS containing ampicillin and streptomycin sulphate to be rinsed, and the rinsed human autogenous abdominal skin is added into a DMEM containing collagenase and hyaluronidase and is placed overnight; PBS digestive juice containing pancreatin and EDTA is added, cells are collected in a centrifugal mode, and the collected cells are eluted with a DMEM and then cultured in an open mode; the cells are subcultured in a certain proportion and prepared into cell suspension; the cell suspension is taken and transferred into a centrifugal tube, and autogenous skin fibroblasts are extracted in a centrifugal mode; various factors are added into the autogenous skin fibroblasts, and a mixture is obtained and applied to beautifying and anti-aging. According to the method, the biological effects of the autogenous skin fibroblasts are utilized, the nutritional factors are added, and the mixture is better stored and used in time within the biological shelf life and can perform the optimal beautifying effects such as repair, regeneration, filling, freckle removal and oxidation resistance on the skin.

Description

technical field [0001] The invention belongs to the technical field of biological cells, and relates to a preparation method of autologous skin fibroblasts for beauty and anti-aging. Background technique [0002] At present, the most commonly used target cells for somatic cell gene therapy are: bone marrow cells, fibroblasts, keratinocytes, liver cells, endothelial cells, muscle cells, etc. Fibroblasts are the main cell components of the skin and have the ability to synthesize and secrete three types of fibers APSC The function of pluripotent cells and matrix plays a very important role in the process of interstitial renewal and wound repair. Fibroblasts have become an ideal target cell due to their many advantages. The skin is the largest organ in the human body. Fibroblasts are easier to obtain, culture, passage and cryopreservation, which makes it possible for fibroblasts to be used in gene therapy. [0003] With the improvement of people's living standards, people pay m...

Claims

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Application Information

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IPC IPC(8): A61K8/99A61K8/98A61K8/44A61K8/64A61K8/65A61K8/67A61Q19/08C12N5/071
CPCA61K8/44A61K8/64A61K8/65A61K8/673A61K8/985A61K8/99A61Q19/08C12N5/0656
Inventor 张正亮
Owner ANHUI HUIEN BIOTECH
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