Freezing resuscitation solution for multi-cytokine-induced killer cells, and applications thereof

A technology for killing cells and cytokines, which is applied in the field of bioengineering and can solve problems such as the clinical application of inappropriate cell products

Inactive Publication Date: 2018-07-06
SOUTH CHINA INSTITUDE OF BIOMEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since fetal bovine serum is of animal origin, it is not suitable for clinical application of cell products

Method used

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  • Freezing resuscitation solution for multi-cytokine-induced killer cells, and applications thereof
  • Freezing resuscitation solution for multi-cytokine-induced killer cells, and applications thereof
  • Freezing resuscitation solution for multi-cytokine-induced killer cells, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] 1. PBMC recovery

[0053] (1) Peripheral blood mononuclear cells (PBMCs) cryopreserved in liquid nitrogen were taken out and rapidly thawed in a water bath at 37-42°C.

[0054] (2) The cells were treated with an equal volume of X-VIVO15 medium (referred to as group P) and an equal volume of recovery solution containing 2.5% HSA and 5% Dextran40 (referred to as group F1).

[0055] (3) Centrifuge, take the cells separately, count with a cell viability analyzer, and measure the cell viability.

[0056] 2. Induced into CIK cells

[0057] Cells in group P and group F1 were cultured statically for 24 hours in X-VIVO15 medium, respectively. Then, 1000U / ml IFN-γ and 2.5% Supgrow were added to the cell culture medium of the P group and the F1 group, after continuing to culture for 24 hours, 100ng / ml CD3, 1000U / ml IL-2, 1000U / ml IL-1α were added, Perform in vitro induction amplification.

[0058] 3. Calculate

[0059] Since the number of initial cells is inconsistent when re...

Embodiment 2

[0086] 1. Cord blood MNC resuscitation

[0087] (1) The cryopreserved umbilical cord blood mononuclear cells (MNC) were taken out from liquid nitrogen and rapidly thawed in a water bath at 37-42°C.

[0088] (2) Use equal volumes of X-VIVO15 medium (referred to as P group), and equal volumes of resuscitation fluid containing 2.5% HSA and 5% Dextran40 (referred to as F1 group), 2.5% plasma, 5% Dextran40 Cells were treated in four ways with resuscitation solution (referred to as F2 group), resuscitation solution with 5% plasma and 5% Dextran40 (referred to as F3 group).

[0089] (3) Centrifuge, take the cells separately, and use a cell viability analyzer to count and measure the cell viability.

[0090] 2. Induced into CIK cells

[0091] The cells treated in groups P and F1-3 were cultured statically for 24 hours in X-VIVO15 medium, respectively. The cells treated in the P and F1-3 groups were treated by adding 1000U / ml IFN-γ and 2.5% Supgrow, and continued to culture for 24 h...

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PUM

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Abstract

The invention discloses a freezing resuscitation solution for multi-cytokine-induced killer cells, and applications thereof, wherein the freezing resuscitation solution comprises albumin and dextran.According to the present invention, the multi-cytokine-induced killer cells obtained through the induction and amplification of the mononuclear cells resuscitated with the freezing resuscitation solution have advantages of rapid proliferation, high induction differentiation efficiency and good cell viability.

Description

technical field [0001] The present invention relates to the field of bioengineering, in particular, the frozen resuscitation liquid for killing cells induced by various cytokines and its application, more specifically, the frozen resuscitation liquid for killing cells induced by various cytokines and the Kit for freezing and thawing of various cytokine-induced killer cells. Background technique [0002] Cytokine-induced killer cells (CIK) are a group of heterogeneous cells obtained from mononuclear cells in human blood induced and expanded by various cytokines in vitro. The cells express both CD3+ and CD56+ cell membrane surface markers, so they are called NK cell-like T lymphocytes, which have T lymphocyte activity and non-MHC restriction of NK cells. Therefore, it can be used to activate the body's immune system and improve the body's immunity; CIK cells can secrete various cytokines such as IL-2, IL-6, IFN-γ, and treat leukemia, lymphoma, lung cancer, gastric cancer, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783A01N1/02
Inventor 裴雪涛陈琳陈海明岳文姚海雷习佳飞何丽娟张博文贾雅丽南雪谢小燕
Owner SOUTH CHINA INSTITUDE OF BIOMEDICINE
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