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

Mesenchymal stem cell cryopreservation medium, cryopreservation method, preservation kit and recovery method

A technology for stem cells and preservation reagents, applied in the field of cell biology, can solve the problems of high DMSO content, easy to be unevenly heated, and high cytotoxicity, and achieve the effect of reasonable cell density, protection of cell activity, and maintenance of cell activity.

Active Publication Date: 2021-12-17
GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
View PDF9 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, animal serum is usually used in this cell cryopreservation solution, which will produce exogenous contamination from xenobiotic sources, high DMSO content, and high cytotoxicity. This cell cryopreservation solution cannot be used clinically
Moreover, the general cell recovery process is prone to uneven heating, secondary crystallization, and greater damage to cells.

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
  • Mesenchymal stem cell cryopreservation medium, cryopreservation method, preservation kit and recovery method
  • Mesenchymal stem cell cryopreservation medium, cryopreservation method, preservation kit and recovery method
  • Mesenchymal stem cell cryopreservation medium, cryopreservation method, preservation kit and recovery method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] A method for cryopreserving mesenchymal stem cells, comprising the following steps:

[0045] Digestion: Take out the mesenchymal stem cells with a confluence of 80%, wash them once with normal saline, add an appropriate amount of 0.25% trypsin to digest them, add selective medium to stop the digestion after the cells become round, centrifuge at 300g for 5min, and retain the precipitate. Add the mesenchymal stem cell cryopreservation solution, resuspend the cell solution, and control the cell concentration to 5×10 6 pc / mL; Among them, the mesenchymal stem cell cryopreservation medium uses the mesenchymal stem cell selective medium as the base fluid, and each 100mL mesenchymal stem cell cryopreservation medium includes the following components: 30mL mesenchymal stem cell culture supernatant , 5mL glycerol, 5g PVP, 0.5g PVA, 0.5g trehalose, 1g HA nanoparticles, 1mg all-trans retinoic acid, 10mL serum replacement; the mesenchymal stem cell culture supernatant was obtained b...

Embodiment 2

[0051] A mesenchymal stem cell cryopreservation method, which differs from the cryopreservation method in Example 1 in that each 100mL mesenchymal stem cell cryopreservation solution includes the following components: 30mL mesenchymal stem cell culture supernatant, 5mL glycerol, 5g PVP, 0.5g PVA, 0.5g trehalose, 2g HA nanoparticles, 1mg all-trans retinoic acid, 10mL serum replacement.

[0052] The recovery method of the cryopreserved cells is the same as the recovery method in Example 1.

Embodiment 3

[0054] A cryopreservation method for mesenchymal stem cells, which is the same as the cryopreservation method in Example 1.

[0055] The resuscitation method for cryopreserved cells is different from the resuscitation method in Example 1 in that each 100 mL of resuscitation solution includes the following components: 0.3 g of glucose, 1 mg of all-trans retinoic acid, and 1 mg of ascorbic acid.

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 mesenchymal stem cell cryopreservation solution, a cryopreservation method, a preservation kit and a recovery method, and relates to the technical field of cell biology. The mesenchymal stem cell cryopreservation solution includes the following components at working concentrations: mesenchymal stem cell culture supernatant 25-35mL / 100mL, glycerin 4-6mL / 100mL, polyvinylpyrrolidone 4-6g / 100mL, polyvinyl alcohol 0.5~1.5g / 100mL, trehalose 0.5~2.5g / 100mL, hydroxyapatite nanoparticles 1~2.5g / 100mL, all-trans retinoic acid 0.5~1.5mg / 100mL, serum substitute 8~12mL / 100mL; The mesenchymal stem cell culture supernatant is prepared by the following method: mesenchymal stem cells are cultured at 78-82% confluence for 22-26 hours, filtered, and the filtrate is retained to obtain the mesenchymal stem cell culture supernatant. The cryopreservation solution of the invention can effectively reduce the damage of cells in the process of cryopreservation and resuscitation, and improve the activity of cells after resuscitation.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a mesenchymal stem cell cryopreservation solution, a cryopreservation method, a preservation kit and a recovery method. Background technique [0002] Cryoprotectant refers to a substance that can protect cells from frostbite in an extremely low temperature environment, also known as cell cryopreservation solution. Adding cryoprotectants to cell suspensions can protect cells from solution damage and ice crystal damage. The cryoprotectant combines with the water molecules in the solution to cause a hydration reaction, weakens the crystallization of water, increases the viscosity of the solution to reduce the formation of ice crystals, and at the same time maintains a certain molar concentration inside and outside the cells to reduce the concentration of unfrozen solutions inside and outside the cells. The concentration of electrolytes in the medium protects cells from damage ...

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): A01N1/02C12N5/0775
CPCA01N1/0221A01N1/0226C12N5/0662
Inventor 孙灿兴李静静赵蓝刘小翠江嘉豪
Owner GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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