A kind of immune cell serum-free cryopreservation liquid and cryopreservation method
A technology of immune cells and cryopreservation solution, which is applied in the field of cell culture, can solve the problems of changing cell thermodynamics, chemical and physical environment, single cell damage, biological damage, etc., and achieve the goal of improving survival rate, good proliferation ability and good activity Effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
preparation example Construction
[0025] The preparation process of NK cells in Examples 1 to 3 is as follows:
[0026] (1) Take an appropriate amount of blood, divide it evenly into two 50ml centrifuge tubes, centrifuge at 650g at room temperature (both reduce the lifting speed to 1), and centrifuge for 15min;
[0027] (2) Remove the upper layer of plasma after centrifugation, add an equal volume of PBS to the lower layer of cells and mix to obtain a cell suspension;
[0028] (3) Carefully add to the Ficoll layer to keep the layers clear, room temperature, and centrifuge;
[0029] (4) Aspirate the peripheral blood mononuclear cell (PBMC) layer, aspirate the cell layer at the junction of the two liquid surfaces as much as possible, add PBS to mix, centrifuge, and wash the cells again in the same way to obtain peripheral blood mononuclear cells (PBMC);
[0030] (5) The above-mentioned PBMC cells were screened with magnetic beads to obtain NK cells.
[0031] The present invention has no special limitation on t...
Embodiment 1
[0033] A serum-free cryopreservation solution for NK cells, including serum-free RPMI 1640 medium and a cell cryopreservation agent, the volume fraction of the cell cryopreservation agent in the serum-free cryopreservation solution for immune cells is: 8%. Based on the final volume of the stock solution, the content of each component in the cell protection agent is: polyethylene glycol 4.0 mg / mL, nonylphenol polyoxyethylene ether-10 2.0 mg / mL, saponins 8.5 μg / mL, Sodium carboxymethyl cellulose 10.5 μg / mL, polyglycerol fatty acid ester 16.0 μg / mL, bamboo leaf flavone 38.0 μg / mL.
[0034] A method for freezing NK cells. The NK cells to be frozen are mixed with the above-mentioned serum-free cell freezing solution to obtain a cell suspension, and the concentration of cells in the cell suspension is 5×10 6 cells / mL, the above cell suspension was placed in a sterile cryopreservation tube and cooled to -80°C, frozen for 12 hours, and then frozen in liquid nitrogen.
Embodiment 2
[0036] A serum-free cryopreservation solution for NK cells, including serum-free RPMI 1640 medium and a cell cryopreservation protection agent, the volume fraction of the cell cryopreservation protection agent in the serum-free cryopreservation solution for immune cells is: 5%. Based on the final volume of the stock solution, the content of each component in the cell protection agent is: polyethylene glycol 3.5 mg / mL, nonylphenol ethoxylate-10 1.5 mg / mL, saponins polysaccharide 8.5 μg / mL, Sodium carboxymethyl cellulose 9.0 μg / mL, polyglycerol fatty acid ester 15.0 μg / mL, bamboo leaf flavone 35.0 μg / mL.
[0037] A method for freezing NK cells. The NK cells to be frozen are mixed with the above-mentioned serum-free cell freezing solution to obtain a cell suspension. The concentration of cells in the cell suspension is 9×10 6 cells / mL, the above cell suspension was placed in a sterile cryopreservation tube and cooled to -80°C, frozen for 20 hours, and then frozen in liquid nitrog...
PUM
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