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

Lymphocyte cryopreservation method

A cryopreservation method and lymphocyte technology, applied in the biological field, can solve the problems of changing cell thermodynamics, chemical and physical environment, biological damage, etc., so as to reduce immune response, reduce cell damage, and reduce low temperature damage to cells. Effect

Inactive Publication Date: 2017-10-31
北京弘润天源基因生物技术有限公司
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cryopreservation process will significantly change the thermodynamic, chemical and physical environment of cells, accompanied by the risk of biological damage

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1) adding the separation liquid containing trehalose, ultra-low viscosity sodium alginate and polydiallyldimethylammonium chloride into the centrifuge tube;

[0027] 2) Add an equal volume of normal saline or serum-free medium to the anticoagulant-treated peripheral blood or umbilical cord blood, dilute it and spread it on the separation medium, at 8-25°C, 200g-800g, 10-30min Centrifuge, absorb the upper layer of plasma for later use, and absorb buffy coat cells to obtain lymphocytes;

[0028] 3) Preparation of cryopreservation solution: 12% DMSO + 10% dextran + 18% upper plasma + 1% trehalose + 59% saline for injection were mixed as a cryopreservation solution;

[0029] 4) Mix the lymphocytes and cryopreservation solution at a ratio of 0.9 to 1.1:1, and distribute them in cell cryopreservation tubes;

[0030] 5) The cell cryopreservation tubes were placed in the programmed cryopreservation box, frozen at -80°C, and then transferred to liquid nitrogen for cryopreservat...

Embodiment 2

[0033]1) adding the separation liquid containing trehalose, ultra-low viscosity sodium alginate and polydiallyldimethylammonium chloride into the centrifuge tube;

[0034] 2) Add an equal volume of normal saline or serum-free medium to the anticoagulant-treated peripheral blood or umbilical cord blood, dilute it and spread it on the separation medium, at 8-25°C, 200g-800g, 10-30min Centrifuge, absorb the upper layer of plasma for later use, and absorb buffy coat cells to obtain lymphocytes;

[0035] 3) Preparation of cryopreservation solution: 12% DMSO + 10% dextran + 18% upper plasma + 1% trehalose + 59% saline for injection were mixed as a cryopreservation solution;

[0036] 4) Mix the lymphocytes and cryopreservation solution at a ratio of 0.9 to 1.1:1, and distribute them in cell cryopreservation tubes;

[0037] 5) The cell cryopreservation tubes were placed in the programmed cryopreservation box, frozen at -80°C, and then transferred to liquid nitrogen for cryopreservati...

Embodiment 3

[0040] 1) adding the separation liquid containing trehalose, ultra-low viscosity sodium alginate and polydiallyldimethylammonium chloride into the centrifuge tube;

[0041] 2) Add an equal volume of normal saline or serum-free medium to the anticoagulant-treated peripheral blood or umbilical cord blood, dilute it and spread it on the separation medium, at 8-25°C, 200g-800g, 10-30min Centrifuge, absorb the upper layer of plasma for later use, and absorb buffy coat cells to obtain lymphocytes;

[0042] 3) Preparation of cryopreservation solution: 12% DMSO + 10% dextran + 18% upper plasma + 1% trehalose + 59% saline for injection were mixed as a cryopreservation solution;

[0043] 4) Mix the lymphocytes and cryopreservation solution at a ratio of 0.9 to 1.1:1, and distribute them in cell cryopreservation tubes;

[0044] 5) The cell cryopreservation tubes were placed in the programmed cryopreservation box, frozen at -80°C, and then transferred to liquid nitrogen for cryopreservat...

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 discloses a cryopreservation method for lymphocyte and belongs to the technical field of biology. The cryopreservation method includes the following steps that a separating medium containing mycose, ultra-low viscosity sodium alginate and poly dimethyl diallyl ammonium chloride is added to a centrifugal tube; after normal saline or a serum-free medium with the same volume as peripheral blood or umbilical cord blood is added to the peripheral blood or the umbilical cord blood treated with anticoagulant for dilution, the mixture is flatly laid on the separating medium, centrifugation is conducted, upper-layer plasma is sucked for use, albuginea layer cells are sucked, and the lymphocyte is obtained; cryopreservation liquid is prepared, wherein 12% of DMSO, 10% of dextranum, 18% of upper-layer plasma, 1% of trehalose and 59% of injection normal saline are mixed to serve as the cryopreservation liquid; the lymphocyte and the cryopreservation liquid are evenly mixed according to the proportion of (0.9-1.1):1 and sub-packaged in a cell cryopreservation tube. The components of the separating medium are partially similar to those of the cryopreservation liquid, damage of allogenic materials to the cells is reduced, and safety is improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a freezing method for lymphocytes separated from peripheral blood or umbilical cord blood. Background technique [0002] As a kind of effector cells, lymphocytes widely exist in human peripheral blood, bone marrow or human umbilical cord blood. The acquisition of high-purity lymphocytes is of great significance for the cytological diagnosis of diseases, component blood transfusion, and the preparation of therapeutic cell products. In the course of clinical application, a common problem is that it is difficult to ensure a sufficient number of lymphocytes. To solve this problem, the current common practice is to use cryopreservation technology to preserve peripheral blood or umbilical cord blood cells, revive and induce culture into various immune cells when needed. However, the cryopreservation process will significantly change the thermodynamic, chemical and physical environment of...

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): A01N1/02
CPCA01N1/02A01N1/021A01N1/0221A01N1/0226A01N1/0284
Inventor 宫喜魁李若鲲马艳马艳玲郝丽敏
Owner 北京弘润天源基因生物技术有限公司
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