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Cell freezing medium

A technology for cryopreservation and cells, which is applied in the field of cell therapy and can solve the problems of introduction of pollution and poor cell activity.

Active Publication Date: 2014-02-12
BEIJING YONGTAI IMMUNITY APPL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cell cryopreservation fluids commonly used in laboratories or commercially available cell freezing fluids usually need to be added with xenogeneic serum or xenogeneic proteins, so there is a risk of introducing contamination and allergens when used for immune cell therapy
However, the cell cryopreservation solution clinically used for cryopreservation of umbilical cord blood stem cells has the disadvantage of poor cell activity after resuscitation when cryopreserving immune cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1, the comparison of three kinds of different cell cryopreservation liquids

[0018] 1. Experimental Design

[0019] Taking the commonly used cell freezing solution in the laboratory (fetal bovine serum+DMSO, hereinafter referred to as the control freezing solution A; see Situ Zhenqiang et al. "Cell Culture") as a contrast, compare the cell freezing solution developed by the inventors (cell freezing Preservation solution C) and umbilical cord blood hematopoietic stem cells commonly used cryopreservation solution (Dextran-40+DMSO, hereinafter referred to as umbilical cord blood cryopreservation solution B; Rubinstein et al., Proc.Nati.Acad.Sci.USA, Vol.92, pp.10119 -10122, Oct1995) for cryopreservation performance of twice subcultured lymphocytes.

[0020] 2. Experimental method

[0021] 2.1 Isolation and inoculation of peripheral blood mononuclear cells

[0022] Peripheral blood was collected from 3 healthy donors, 45 mL / person, and peripheral blood mononu...

Embodiment 2

[0044] Embodiment 2, to the optimization of cell cryopreservation liquid formula

[0045] 1 Experimental design

[0046] Adjust the composition of each component in the cryopreservation solution C formula in Example 1 (Table 3 below), and find the optimal concentration ratio through experimental comparison. Experimental method is the same as embodiment 1.

[0047]Table 3 optimizes the cryopreservation solution C

[0048] group

20%HSA(v / v)

DMSO (v / v)

Dextran 40 Glucose Injection (v / v)

1

10％

5％

85％

2

20％

5％

75％

3

30％

5％

65％

4

40％

5％

55％

[0049] 2 Experimental results

[0050] The experimental results are shown in Table 4 below. Wherein the second group of experimental results (i.e. the cryopreservation solution C in Example 1) is the best, but other groups are also better than cord blood cryopreservation solution B.

[0051] Table 4. Experimental results of modified cryo...

Embodiment 3

[0053] Embodiment 3, to the cryopreservation effect of the cell of different concentration

[0054] 1. Experimental Design

[0055] The cryopreservation solution C prepared in Example 1 was used to freeze cells with different concentrations, and the number and activity of cells after thawing were detected, and the applicable cell concentration range of the cryopreservation solution C was studied.

[0056] 2. Experimental method

[0057] Operations such as cell subculture, recovery after cryopreservation, etc. were as in Example 1. Prepare 50 ml of cell freezing solution according to the formula of freezing solution C in Example 1. Take the cell culture bottle out of the cell culture incubator, pat and mix the cell suspension in the bottle, and use a pipette to draw 0.5 mL for cell counting. According to the counting results, divide into three cell suspensions, and refer to the following table for the number of cells in each portion:

[0058] Table 5. Different cell freezin...

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Abstract

The invention provides a cell freezing medium which can be clinically safely used and is used for freezing human cells. The cell freezing medium comprises: 2-8wt% of human albumin, 5-10 volume% of dimethyl sulfoxide, 3.5-5.1wt% of dextranum-40 and 2.75-4.25wt% of glucose.

Description

field of invention [0001] The invention relates to the field of cell therapy, in particular to a cell cryopreservation solution that can be safely used in clinical practice. Background of the invention [0002] Immune cell therapy has gradually become one of the important adjuvant methods in the routine clinical treatment of tumors and is widely used in clinical practice. In the process of immune cell preparation or clinical application, under certain circumstances, cells need to be cryopreserved in liquid nitrogen, and then used for further culture or clinical direct reinfusion after a period of time. The cell cryopreservation solution commonly used in the laboratory or the commercially available cell cryopreservation solution usually needs to be added with xenogeneic serum or protein, so there is a risk of introducing contamination and allergens when used for immune cell therapy. However, the cell cryopreservation solution clinically used for cryopreservation of umbilical...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0226
Inventor 邵谊
Owner BEIJING YONGTAI IMMUNITY APPL TECH
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