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

A method for expanding umbilical cord mesenchymal stem cells and its application in arthritis

A technology of mesenchymal stem cells and umbilical cords, applied in the field of osteoarthritis treatment, can solve the problems of limited expansion capacity and achieve the effect of fast expansion

Active Publication Date: 2021-08-27
杨凌洛威塔生物科技有限责任公司
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a method for the expansion of umbilical cord mesenchymal stem cells to solve the problems of limited expansion ability in the cultivation of umbilical cord mesenchymal stem cells in the actual application process, and to use the mesenchymal stem cells combined with other components Joint treatment of osteoarthritis for better treatment results

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
  • A method for expanding umbilical cord mesenchymal stem cells and its application in arthritis
  • A method for expanding umbilical cord mesenchymal stem cells and its application in arthritis
  • A method for expanding umbilical cord mesenchymal stem cells and its application in arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Isolation and culture of mesenchymal stem cells. In the operating room, the umbilical cord is removed, stored in sterile saline, and transported to the laboratory on a cold chain. On the 100-level ultra-clean workbench of the GMP laboratory, use sterile surgical instruments to peel off Wharton's jelly, add it to 10 times the volume of trypsin digestion solution (0.5% trypsin + 0.2% EDTA), and keep at 37 degrees Lower digestion 2h. Then add 1mg / mL type I collagen and continue to digest for 1h. Pass the above suspension through a 200-mesh sieve, blow and filter. The above cell suspension was mixed with the culture medium, resuspended, added to a T25 cell culture flask, and cultured in a 37°C, 5% carbon dioxide incubator until generation P1.

Embodiment 2

[0041] Example 2: Divide the experiment into five groups, Group A (DMEM / F12+ 10%PRP+5ng / mL bFGF), Group B (DMEM / F12+ 10%PRP+10ng / mL BFGF), Group C (DMEM / F12+ 10 %PRP + 20ng / mL bFGF), group D (DMEM / F12+10%PRP + 50ng / mL bFGF), group E (DMEM / F12+ 10%PRP + 100ng / mL bFGF).

[0042] After centrifugation, the P1 generation cells were resuspended in DMEM / F12 medium containing 10% PRP, and the density was adjusted to 5×10 4 cells / mL, inoculate five 24-well plates at 500 μL / well. According to the grouping, in the corresponding wells, add different concentrations of bFGF, 24 wells in each group. Subsequently, five 24-well plates were placed in a 37-degree, 5% carbon dioxide incubator for cultivation, and were cultured at 1 d, 2 d, 3 d, 4 d, 5 d, 6 d, 7 d, 8 d, a total of 8 time points , cells from 3 wells in each group were taken at each time point for cell counting.

[0043] figure 1 The experimental results showed that the cells in group D proliferated the fastest. It shows that t...

Embodiment 3

[0044] Example 3: Divide the experiment into five groups, group A (DMEM / F12+ 10%PRP + 20U / mL PDGF), group B (DMEM / F12+ 10%PRP + 50U / mL PDGF), group C (DMEM / F12+ 10 %PRP + 100U / mL PDGF), group D (DMEM / F12+10%PRP + 200U / mL PDGF), group E (DMEM / F12+ 10%PRP + 500U / mL PDGF).

[0045] After centrifugation, the P1 generation cells were resuspended in DMEM / F12 medium containing 10% PRP, and the density was adjusted to 5×10 4 cells / mL, inoculate five 24-well plates at 500 μL / well. According to the grouping, different concentrations of PDGF were added to the corresponding wells, 24 wells in each group. Subsequently, five 24-well plates were placed in a 37-degree, 5% carbon dioxide incubator for cultivation, and were cultured at 1 d, 2 d, 3 d, 4 d, 5 d, 6 d, 7 d, 8 d, a total of 8 time points , cells from 3 wells in each group were taken at each time point for cell counting.

[0046] figure 2 The experimental results showed that the cell proliferation in group C was the fastest. It...

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 present invention relates to the improvement of the separation and culture technology of umbilical cord mesenchymal stem cells and its application in arthritis. The present invention provides a method of separation and culture of umbilical cord-derived mesenchymal stem cells and its application in arthritis, which includes the following steps: 1) Mesenchymal stem cells were obtained from human umbilical cord by enzymatic digestion; 2) Mesenchymal stem cells were cultured with DMEM / F12 medium supplemented with basic fibroblast growth factor and platelet-derived growth factor combined with platelet-rich plasma; 3) Improvement of hyaluronic acid, adding methacrylic anhydride to hyaluronic acid to form a 3D bio-scaffold, which can fix mesenchymal stem cells on bone and joint injuries for fixed-point repair; 4) cultured mesenchymal stem cells , combined with PRP and modified hyaluronic acid, injected into the bones and joints of volunteers together to treat osteoarthritis.

Description

technical field [0001] The invention relates to a method for expanding umbilical cord mesenchymal stem cells, and using the mesenchymal stem cells in combination with other components to treat osteoarthritis. Background technique [0002] Osteoarthritis is a progressive and painful disease that can affect both the young and the elderly. In an aging society, the prevalence is likely to increase further. It is estimated that at least 100 million people nationwide are affected by arthritis. Worldwide, arthritis is the fourth leading cause of disability. Osteoarthritis is an important cause of disability in both developed and developing countries. [0003] Osteoarthritis is a progressive and irreversible degenerative disease of cartilage. Articular cartilage is very poorly repairable and avascular, so lack of systemic regulation may lead to ineffective healing. There is evidence that osteoarthritis is associated with depletion of stromal mesenchymal stem cells, manifested b...

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 Patents(China)
IPC IPC(8): C12N5/0775A61K35/28A61K35/16A61K47/36A61P19/02A61P29/00C08B37/08
CPCA61K9/0019A61K35/16A61K35/28A61K47/36A61P19/02A61P29/00C08B37/0072C12N5/0668C12N2501/115C12N2501/135C12N2509/00A61K2300/00
Inventor 刘建强何晓艾霞
Owner 杨凌洛威塔生物科技有限责任公司
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