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Blood preservation solution

A technology of blood preservation solution and preservation solution, which is applied in the field of cell therapy and can solve the problems of verification of the subsequent proliferation ability of lymphocytes.

Active Publication Date: 2016-08-17
BEIJING YONGTAI IMMUNITY APPL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patent application CN 1411818A discloses an anticoagulant for lymphocytes in peripheral blood, but this application does not verify the subsequent proliferation of lymphocytes preserved by using the disclosed anticoagulant

Method used

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Experimental program
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Effect test

Embodiment 1

[0033] Embodiment 1, three kinds of anticoagulants / preservation solution performance comparison

[0034] 1. Preservation solution preparation

[0035] The heparin anticoagulant (herein referred to as H) is a lithium heparin vacuum anticoagulant tube (Greiner, 9ml).

[0036] The formula of the anticoagulant disclosed in CN 1411818A (hereinafter referred to as S) is as follows in Table 1:

[0037] Formulation S in Table 1: per 100ml

[0038]

[0039] The specific preparation method is as follows (20ml): first dissolve 664mg of trisodium citrate, 5mg of citric acid and 550mg of glucose for injection in 12ml. Then dilute to 20ml with water for injection. Adjust the pH to 6.8-7.3 using NaOH and HCl. Filter with a 0.22um needle filter, then aliquot 5ml / tube, and store at 4°C.

[0040] Preservation solution of the present invention (herein referred to as N) formula is as follows table 2:

[0041] Formulation N in Table 2: per 100ml

[0042]

[0043]The specific preparati...

Embodiment 2

[0068] Embodiment 2, preservation solution formulation optimization

[0069] On the basis of Example 1, the inventors studied the amount of SOD in formula N, so as to optimize the protective performance of the preservation solution on lymphocytes.

[0070] Peripheral blood (54ml / person) was collected from three healthy donors, and the blood samples were stored at 2-8°C in preservation solutions containing three concentrations of SOD (70U / 100ml, 280U / 100ml, 560U / 100ml) at a ratio of 1 : 10 (preservation solution: peripheral blood). 3ml of peripheral blood was taken out at five time points of 0h, 12h, 24h, 36h, and 48h, and peripheral blood mononuclear cells (PBMCs) were separated by Ficoll-Hypaque (GE) density gradient centrifugation, and samples were taken for detection. Cell culture medium (IMSF100, Immunotech UK.Ltd) was resuspended and inoculated into culture flasks coated with activated antibodies (Ebioscience) for expansion culture. The cell culture medium was replenish...

Embodiment 3

[0091] Embodiment 3, optimization of preservation solution usage ratio

[0092] On the basis of the foregoing examples, the inventors studied the proportion of formula N used so as to optimize the protective performance of the preservation solution on lymphocytes.

[0093]Collect peripheral blood (54ml / person) from three healthy donors, mix with the preservation solution of the present invention in three proportions respectively (preservation solution: blood sample=1:20, 1:10, 1:5), and store the blood sample at 2-8 ℃. 3ml of peripheral blood was taken out at five time points of 0h, 12h, 24h, 36h, and 48h, and peripheral blood mononuclear cells (PBMCs) were separated by Ficoll-Hypaque (GE) density gradient centrifugation, and samples were taken for detection. Cell culture medium (IMSF100, Immunotech UK.Ltd) was resuspended and inoculated into culture flasks coated with activated antibodies (Ebioscience) for expansion culture. The cell culture medium was replenished once at 7...

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Abstract

The invention provides a new blood preservation solution. One hundred milliliters of the blood preservation solution contain 3.31-3.32g of sodium citrate, 0.01-0.05g of citric acid, 2.45-3.37g of glucose, 0.2-0.4g of ATP, 50-600U of SOD, and the balance of water. The present invention also provides a method for preserving a blood sample by using the blood preservation solution.

Description

technical field [0001] The invention relates to the field of cell therapy, in particular to a long-acting blood preservation solution for maintaining the expansion effect of peripheral blood lymphocytes and a method for using the same. Background of the invention [0002] Malignant tumor is one of the main diseases that threaten human health and life, and its incidence rate is increasing year by year. In the 1980s, Rosenberg et al. observed that after animals with non-immune system tumors were treated with recombinant IL-2, the animal's lymphocytes could regulate tumor regression and metastasis. Since then, the clinical application of immune cell therapy has rapidly expanded, and it has become the fourth mode of comprehensive tumor treatment besides surgery, chemotherapy and radiotherapy. Peripheral venous blood is the raw material for culturing cells for all immune cell therapies, and heparin or compound sodium citrate preservation solution (ACD) is generally added to veno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 邵谊
Owner BEIJING YONGTAI IMMUNITY APPL TECH
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