Method for simultaneously preparing autologous epidermal cells and fibroblasts, and biological beauty product comprising autologous epidermal cells and fibroblasts
A technology of fibroblasts and epidermal cells, applied in animal cells, cosmetic preparations, cosmetic preparations, etc., can solve the problems of increased usage, insufficient skin beauty efficacy, poor effect, etc., and achieves regeneration and proliferation. Effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0049] Example 1 Preparation of human autologous epidermal cells
[0050] 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.
[0051] 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 0.25% (mass volume ratio) trypsin to digest for 30 minutes, wherein trypsin contains mass volume ratio 0.01% EDTA; pass through a 40 μm cell sieve after digestion, add 10ml 1×PBS after filtration, cent...
Embodiment 2
[0052] Example 2 Preparation of human autologous fibroblasts
[0053] 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 0.25% trypsin to digest for 30 minutes, pass through a 40μm cell sieve after digestion, add 10ml 1 ×PBS, centrifuge at 1000 rpm for 10 minutes to obtain a single cell pellet, resuspend in 10ml 1×PBS again, and centrifuge at 1000 rpm for 10 minutes to obtain a single cell pellet, add α-MEM medium to resuspend, Counting, using 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 fresh culture medium every 2 d...
Embodiment 3
[0055] Example 3 Detection of specific protein expression in human autologous epidermal cells and fibroblasts
[0056] 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.
[0057] According to the molecular weight of the target protein, a 10% separating gel was prepa...
PUM
Property | Measurement | Unit |
---|---|---|
greyscale | aaaaa | aaaaa |
greyscale | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com