Method for preserving and transporting immune cells

A transportation method and technology of immune cells, applied in the field of storage and transportation of immune cells, can solve the problems of single cell damage, unfavorable clinical application, changes in cell thermodynamics, chemical and physical environment, etc., to reduce cell damage and raw material uncertainty. Less sex, reducing the effect of immune response

Inactive Publication Date: 2016-05-04
北京弘润天源基因生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research and clinical application of immune cells is a major and complex task coordinated by multiple disciplines, units, and departments. It is necessary to have an independent cell preparation unit to provide cells, an independent cell transplantation unit to perform cell transplantation, and a cell preparation unit There is a certain space distance between the unit and the cell transplantation unit. In order to make the cell preparation unit and the cell transplantation unit fully connected in the work, so that the cell transplantation can achieve the best performance, there must be a standardized cell storage and transportation process that can meet the standards. The original immune cells that need to be st

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The method for storing and transporting immune cells comprises the following steps:

[0025] 1) collecting peripheral blood or umbilical cord blood, adding an equal volume of normal saline or serum-free medium to dilute to obtain a cell suspension;

[0026] 2) Add the separation liquid containing trehalose, ultra-low viscosity sodium alginate and amino acid into the centrifuge tube, spread the cell suspension on the separation liquid, at 8-25°C, 200g-800g, 10 Centrifuge under the condition of ~30min, absorb the upper plasma for later use, absorb buffy coat cells to obtain lymphocytes;

[0027] 3) Lymphocytes were mixed in equal proportions with the cryopreservation liquid containing 12% DMSO+18% upper plasma+5% trehalose+5% glycerol+60% normal saline for injection into cell cryopreservation tubes; Seal the mouth of the tube and place it in a freezing box filled with isopropanol; freeze it at -80°C first, and then transfer it to liquid nitrogen for freezing;

[0028] 4) ...

Embodiment 2

[0032] The method for storing and transporting immune cells comprises the following steps:

[0033] 1) collecting peripheral blood or umbilical cord blood, adding an equal volume of normal saline or serum-free medium to dilute to obtain a cell suspension;

[0034] 2) Add the separation liquid containing trehalose, ultra-low viscosity sodium alginate and amino acid into the centrifuge tube, spread the cell suspension on the separation liquid, at 8-25°C, 200g-800g, 10 Centrifuge under the condition of ~30min, absorb the upper plasma for later use, absorb buffy coat cells to obtain lymphocytes;

[0035] 3) Lymphocytes were mixed in equal proportions with the cryopreservation liquid containing 12% DMSO+18% upper plasma+5% trehalose+5% glycerol+60% normal saline for injection into cell cryopreservation tubes; Seal the mouth of the tube and place it in a freezing box filled with isopropanol; freeze it at -80°C first, and then transfer it to liquid nitrogen for freezing;

[0036] 4...

Embodiment 3

[0040] The method for storing and transporting immune cells comprises the following steps:

[0041] 1) collecting peripheral blood or umbilical cord blood, adding an equal volume of normal saline or serum-free medium to dilute to obtain a cell suspension;

[0042] 2) Add the separation liquid containing trehalose, ultra-low viscosity sodium alginate and amino acid into the centrifuge tube, spread the cell suspension on the separation liquid, at 8-25°C, 200g-800g, 10 Centrifuge under the condition of ~30min, absorb the upper plasma for later use, absorb buffy coat cells to obtain lymphocytes;

[0043] 3) Lymphocytes were mixed in equal proportions with the cryopreservation liquid containing 12% DMSO+18% upper plasma+5% trehalose+5% glycerol+60% normal saline for injection into cell cryopreservation tubes; Seal the mouth of the tube and place it in a freezing box filled with isopropanol; freeze it at -80°C first, and then transfer it to liquid nitrogen for freezing;

[0044] 4...

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Abstract

The invention discloses a method for preserving and transporting immune cells and belongs to the technical field of biology. The method comprises the following steps of collecting peripheral blood or umbilical cord blood, and conducting dilution, so that cell suspension is obtained; adding a separating medium containing trehalose, ultra-low-viscosity sodium alginate and amino acid into a centrifuge tube, laying the cell suspension flat on the separating medium, conducting centrifugation, absorbing upper layer blood plasma for use, and absorbing albuginea layer cells, so that lymphocyte is obtained; mixing the lymphocyte with a cryopreservation solution in the same proportion, and placing the mixture into a cell cryopreservation tube; conducting cryopreservation; after the lymphocyte thaws, moving the liquid cell suspension into a centrifuge tube containing normal saline with the temperature ranging from 4 DEG C to 8 DEG C, mixing the mixture uniformly, and conducting centrifugation; adding preserving fluid containing trehalose with the concentration being 2% to the lymphocyte; moving the mixture into a sterile tube or a transfer bag for transfer. According to the method, raw materials meeting the intravenous injection level bulk pharmaceutical chemical standards are selected for all stages from separation to cryopreservation and then to preservation and transportation, uncertainty of the raw materials is low, and safety is high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for storing and transporting immune cells. Background technique [0002] Immune cells not only have good prospects in the treatment of cancer, but also from the perspective of health preservation, immune cells can also be extracted from the peripheral blood of the patient when he is healthy, and the patient himself has immunocompromised or other symptoms in the future When the time comes, the stored immune cells can be reinfused back to the patient to improve the patient's immunity. The research and clinical application of immune cells is a major and complex task coordinated by multiple disciplines, units, and departments. It is necessary to have an independent cell preparation unit to provide cells, an independent cell transplantation unit to perform cell transplantation, and a cell preparation unit There is a certain space distance between the unit and the cell transplant...

Claims

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Application Information

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IPC IPC(8): C12N5/0781C12N5/0783A01N1/02
CPCA01N1/0221A01N1/0236C12N5/0635C12N5/0636
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
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