Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method and kit for efficiently promoting adipose-derived stem cells to proliferate

An adipose stem cell and efficient technology, applied in the direction of cell culture active agent, animal cells, vertebrate cells, etc., can solve the problems of unbearable pain, limited number of adipose stem cells, slow growth rate, etc., to reduce pain, improve medical beauty, Reduce the effect of liposuction again

Pending Publication Date: 2017-07-18
泰州市数康生物科技有限公司
View PDF5 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when adipose stem cells are cultured and proliferated in vitro, the growth rate is slow, reaching 10 8 The above number of cells needs more than 20 days, and it cannot meet the needs of multiple beauty treatments
Many beauticians may need liposuction for the second beauty treatment, but this kind of pain is very painful, and it is difficult for ordinary people to accept
Even with local anesthesia, the pain is unbearable afterwards
[0003] In summary, the currently commonly used adipose stem cell culture method obtains a limited number of adipose stem cells, and the proliferation and growth of adipose stem cells is slow. 8
Moreover, the subculture and expansion of adipose stem cells should not exceed 5 generations, otherwise the adipose stem cells are prone to differentiation and apoptosis, and the functions of cell proliferation and repair are also significantly reduced, and the cells with stem cell characteristics are significantly reduced, and the activity of adipose stem cells after cosmetic treatment cannot be guaranteed.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method and kit for efficiently promoting adipose-derived stem cells to proliferate
  • Method and kit for efficiently promoting adipose-derived stem cells to proliferate
  • Method and kit for efficiently promoting adipose-derived stem cells to proliferate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Preparation of human adipose stem cells

[0059] Cosmetology volunteers obtained 50 mL of adipose tissue from their abdomen or thighs by liposuction (with whom they signed an informed consent form), and washed it with 50 mL of normal saline repeatedly 5 times to remove as much oil and blood cells as possible in the fat. Using 1:1 0.25% (mass volume ratio) trypsin (Beijing Suo Laibao company) and 0.1% (mass volume ratio) type I collagenase (US Sigma company) 50mL, digested in 37 ℃ incubator for 60 minutes, Oscillate with a vortex shaker every 15 minutes for 2 minutes; after digestion, centrifuge at 1500rpm for 10 minutes, absorb the upper layer of fat, add physiological saline to resuspend, and filter with a 100μm cell sieve (BD Company, USA); centrifuge the cell suspension at 1000rpm for 10 minutes, The precipitate was resuspended and washed twice with normal saline, and then added to EGM-2 medium (Lonza Company, USA) (containing 10ng / mL EGF, American R&D Comp...

Embodiment 2

[0062] Example 2 Preparation of gelatin-coated plates and cell culture flasks

[0063] Weigh 5 g of gelatin, dissolve it in 100 mL of 1×PBS (pH=7.4) in a 37° C. water bath, and sterilize it with a 0.45 μm filter to prepare a 5% gelatin solution. Add 500μL gelatin solution to each well of the 6-well plate, add 2mL gelatin solution to the 90mm plate, and add 2mL gelatin solution to the 175cm 2 Add 4 mL of gelatin solution to the culture bottle, incubate overnight at 4°C or 1 hour at 37°C, and then coat in cell culture containers. After coating, the gelatin solution was sucked off, ventilated and dried in a biological safety cabinet or ultra-clean bench, sealed and stored in a 4°C refrigerator for later use.

Embodiment 3

[0064] Example 3 Analysis of Proliferation Activity of Human Adipose Stem Cells

[0065] The adipose stem cells prepared in Example 1 and the adipose stem cells prepared in the comparative example were taken for proliferative activity analysis. The comparative examples were obtained by culturing adipose stem cells according to the article on adipose stem cells in vitro supporting lymphangiogenesis parameters published in the Journal of Cell Biochemistry in November 2016 (Strassburg S, Torio-Padron N, Finkenzeller G, Frankenschmidt A, Stark GB. Adipose-Derived Stem Cells SupportLymphangiogenic Parameters In Vitro.J Cell Biochem.2016Nov;117(11):2620-9.)

[0066] The CCK-8 (Shanghai Beyond Company) kit was used for detection, and the cell proliferation activity was detected on the 1st day, the 2nd day, the 3rd day, the 4th day and the 5th day according to the instruction manual. From figure 1 It can be known that the adipose stem cells prepared in the present invention have fas...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a method for efficiently promoting adipose-derived stem cells to proliferate. The method is characterized in that a cell culture plate or a culture dish coated by gelatin or a cell culture bottle coated by gelatin is adopted to efficiently promote the adipose-derived stem cells to proliferate and grow in fat under combined culture of a recombinant human epithelial growth factor, a recombinant human platelet derived growth factor, a recombinant human basic fibroblast growth factor, insulin, transferrin, fetal calf serum and the like, so that cells from 50mL fat can be cultured to obtain 109 adipose-derived stem cells or more within 15 days, and therefore, great cell population for clinically treating diseases and medical cosmetology can be met, secondary liposuction is avoided and pains caused by liposuction are reduced. According to the method, the invention further provides a kit for efficiently promoting the adipose-derived stem cells to proliferate, and the kit can be used for efficiently promoting the adipose-derived stem cells to proliferate and grow.

Description

technical field [0001] The invention relates to a method for efficiently promoting the proliferation of adipose stem cells and a kit for efficiently promoting the proliferation of adipose stem cells obtained through the method. In particular, the present invention relates to the use of gelatin-coated cell culture plates or dishes and gelatin-coated cell culture flasks in recombinant human epidermal growth factor, recombinant human platelet-derived growth factor, recombinant human basic fibroblast growth factor, insulin Under the joint culture of , transferrin and fetal bovine serum, it can efficiently promote the proliferation and growth of adipose stem cells from fat. The fat stem cells obtained in the invention can be used in medical cosmetology, including the fields of skin wrinkle removal, freckle removal, breast enhancement and the like. Background technique [0002] Due to the rapid development of biotechnology, both medical cosmetology and cosmetics are applied to bi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2533/54C12N2500/25C12N2501/115C12N2501/135C12N2501/11
Inventor 宗玺王锦杰
Owner 泰州市数康生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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