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

Cell cryopreservation liquid and cell cryopreservation method

A cryopreservation method and cryopreservation solution technology, which is applied in the field of cell cryopreservation solution and cell cryopreservation, can solve the problems of high pollution risk and high price, and achieve the effects of low pollution risk, reduced damage, and improved survival rate

Inactive Publication Date: 2014-07-02
SHENZHEN INST OF ADVANCED TECH
View PDF4 Cites 28 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to provide enough nutrition for the cells and maintain the survival rate of the cells, more serum is generally added to the existing cell cryopreservation solution as the nutrient substance for cell growth. etc. are not only expensive, but also easily lead to a higher risk of pollution

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
  • Cell cryopreservation liquid and cell cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] 1. After culturing the 293 cells to the logarithmic phase, in a sterile environment, use an electric pipette to suck up the cell culture medium in the culture dish, add 10mL PBS buffer solution to the culture dish, shake gently to wash, The PBS buffer was then removed with an electric pipette;

[0065] 2. In a sterile environment, evenly add 0.5 mL of 0.25% trypsin solution to the surface of the 293 cells in the culture dish, digest in a 37°C incubator for 2 minutes, and add 3 mL of whole cell culture solution to the culture dish , to terminate the digestion of trypsin, and then use an electric pipette to blow off the 293 cells from the culture dish to suspend the 293 cells in the above-mentioned whole cell culture solution; wherein, the above-mentioned whole cell culture solution includes a volume ratio of 9:1 The cell culture medium and fetal calf serum, the cell culture medium includes the following components:

[0066] Anhydrous calcium chloride 265.00mg / L; ferric ...

Embodiment 2

[0072] 1. After culturing the MCF-7 cells to the logarithmic phase, in a sterile environment, use an electric pipette to suck up the cell culture medium in the culture dish, add 15mL PBS buffer solution to the culture dish, and shake gently. Wash, then remove the PBS buffer with an electric pipette;

[0073] 2. In a sterile environment, evenly add 2 mL of 0.25% trypsin solution to the surface of the MCF-7 cells in the culture dish, digest in a 37°C incubator for 5 minutes, and add 5 mL of whole cell culture to the culture dish solution to stop the digestion of trypsin, and then use an electric pipette to blow off the MCF-7 cells from the culture dish to suspend the MCF-7 cells in the above-mentioned whole cell culture solution; wherein, the above-mentioned whole cell culture solution includes volume Cell culture medium and fetal bovine serum in a ratio of 9:1, the cell culture medium comprising the following components:

[0074] Calcium chloride 116.6mg / L; copper sulfate 0.00...

Embodiment 3

[0080] 1. After culturing HL60 cells to the logarithmic phase, centrifuge the cell culture solution containing the logarithmic phase cells at 800 rpm for 6 minutes, discard the supernatant to obtain the HL60 cell pellet, and then add 12mL PBS buffer to resuspend the cells , then centrifuged at 800 rpm for 6 min, discarded the supernatant to obtain the washed HL60 cell pellet;

[0081] 2. Suspend the washed HL60 cell pellet with cell freezing solution, then count the cells, and dilute with the above cell freezing solution to obtain a concentration of 2×10 6A mixture of HL60 cells in logarithmic phase per milliliter and cell cryopreservation solution; wherein, the above-mentioned cell cryopreservation solution includes 90% whole cell culture solution and 10% dimethyl sulfoxide by volume percentage, and the whole cell The culture medium includes cell culture medium and fetal calf serum with a volume ratio of 9:1, and the cell culture medium includes the following components:

[...

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 relates to a cell cryopreservation liquid and a cell cryopreservation method. The cell cryopreservation liquid comprises 90-95% of a whole cell culture liquid and 5-10% of dimethyl sulfoxide according to the volume percentage, wherein the whole cell culture liquid contains a cell culture medium and fetal calf serum with the volume ratio of 9:1. The whole cell culture liquid of the cell cryopreservation liquid provides essential nutrient substances for cellular life metabolism, and the dimethyl sulfoxide is used as an antifreezing agent; through reasonable proportion of the amounts of the cell culture medium and the fetal calf serum in the whole cell culture liquid and the amounts of the whole cell culture liquid and the antifreezing agent, the whole cell culture liquid with lower content of the fetal calf serum can provide the essential and enough nutrient substances for the cellular life metabolism; moreover, the appropriate amount of the antifreezing agent can prevent or reduce damage effects of frozen ice crystals on cells so as to improve the survival rate of the cells; and the cell cryopreservation liquid contains no serum easily causing pollution, thereby having lower pollution risk.

Description

technical field [0001] The invention relates to cell cryopreservation technology, in particular to a cell cryopreservation solution and a cell cryopreservation method. Background technique [0002] Cell cryopreservation and recovery are one of the important technologies in cell culture technology, and cell cryopreservation is a method to preserve cultured cells. The basic principle of cell cryopreservation and recovery is slow freezing and quick thawing, which can preserve cell viability to the greatest extent. [0003] Cell cryopreservation solution is a solution that must be used for cell cryopreservation. Its function is to suspend cells that need to be frozen in the cryopreservation solution, supply the nutrients necessary for cell life metabolism, and prevent or reduce the impact of frozen ice crystals on cells. damage effect. In order to provide enough nutrition for the cells and maintain the survival rate of the cells, more serum is generally added to the existing c...

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): A01N1/02
Inventor 万晓春陈凤莲赵琦李红昌
Owner SHENZHEN INST OF ADVANCED TECH
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