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Frozen resuscitation solution for multiple cytokine induced killer cells, and application thereof

A technology for killing cells and cytokines, which is applied in the direction of animal cells, vertebrate cells, and cell culture active agents. It can solve the problems of clinical application of unsuitable cell products, and achieve high induction differentiation efficiency, good cell activity, and fast proliferation rate. Effect

Inactive Publication Date: 2018-05-11
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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  • Frozen resuscitation solution for multiple cytokine induced killer cells, and application thereof
  • Frozen resuscitation solution for multiple cytokine induced killer cells, and application thereof
  • Frozen resuscitation solution for multiple cytokine induced killer cells, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] 1. PBMC recovery

[0058] (1) Peripheral blood mononuclear cells (PBMCs) cryopreserved in liquid nitrogen were taken out and rapidly thawed in a water bath at 37-42°C.

[0059] (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).

[0060] (3) Centrifuge, take the cells separately, count with a cell viability analyzer, and measure the cell viability.

[0061] 2. Induced into CIK cells

[0062] 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.

[0063] 3. Calculate

[0064] Since the number of initial cells is inconsistent when re...

Embodiment 2

[0091] 1. Cord blood MNC resuscitation

[0092](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.

[0093] (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).

[0094] (3) Centrifuge, take the cells separately, and use a cell viability analyzer to count and measure the cell viability.

[0095] 2. Induced into CIK cells

[0096] 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 ho...

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Abstract

The invention discloses a frozen resuscitation solution for multiple cytokine induced killer cells, and an application thereof. The frozen resuscitation solution for multiple cytokine induced killer cells comprises albumin and dextran. The multiple cytokine induced killer cells obtained through the subsequent induction and amplification process of single karyocytes obtained through resuscitation using the frozen resuscitation solution have the advantages of fast proliferation rate, high induced differentiation efficiency and good cell viability.

Description

technical field [0001] The present invention relates to the field of bioengineering, in particular, a frozen resuscitation solution for killing cells induced by various cytokines and applications thereof, more specifically, a frozen recovery solution for killing cells induced by various cytokines, used for A kit for cryopreservation of multiple cytokine-induced killer cells, and a method for recovering mononuclear cells and inducing and expanding them into multiple 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 c...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2500/30C12N2501/24C12N2501/998C12N2506/115
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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