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Preparation method for CIK cell with high proliferation and high cell cytotoxic activity

A proliferative and cytotoxic technology, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of low cytotoxic activity and poor proliferation of CIK cells, and achieve the effect of reducing toxic and side effects

Inactive Publication Date: 2007-10-31
HARBIN MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to solve the problems of existing CIK cells with poor proliferative power and low cytotoxic activity, and provide a method for preparing CIK cells with high proliferative power and high cytotoxic activity

Method used

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  • Preparation method for CIK cell with high proliferation and high cell cytotoxic activity
  • Preparation method for CIK cell with high proliferation and high cell cytotoxic activity
  • Preparation method for CIK cell with high proliferation and high cell cytotoxic activity

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specific Embodiment approach 1

[0007] Embodiment 1: In this embodiment, CIK cells with high proliferative power and high cytotoxic activity are prepared according to the following steps: mononuclear cells are suspended in RPMI 1640 medium containing 10% fetal bovine serum for 2 hours, and then recombinant human cytokine IFN is added -γ, so that the concentration of IFN-γ in the suspension is 1000U / mL, after continuing to culture for 24 hours, add CD3 monoclonal antibody protein and gene recombinant human cytokines IL-2, IL-12 and IL-1, so that CD3 monoclonal antibody in the culture medium The concentration of antibody protein is 100ng / mL, the concentration of cytokine IL-2 is 600U / mL, the concentration of cytokine IL-12 is 5ng / mL, and the concentration of cytokine IL-1 is 100U / mL, and then the , CO 2 Concentration of 5% and humidity of 100% continue to culture for 7 to 24 days to obtain CIK cells with high proliferation and high cytotoxic activity; CD3 monoclonal antibody protein and recombinant human cytok...

specific Embodiment approach 2

[0010] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the mononuclear cells are obtained by separating the peripheral blood mononuclear cells from the peripheral blood of healthy adults by Ficoll density gradient separation method. Others are the same as the first embodiment.

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Abstract

The invention discloses a preparing method of high genitality and high cell toxic activity CIK cell, which comprises the following steps: adding gene recombination human cytokine IFN-gamma, CD3 single antibody protein and gene recombination human cytokine IL-2, IL-12 and IL-1 into individual nucleate cell; culturing for 7-24 days under 37 deg. c with CO2 density at 5% and humidity at 100%' getting high genitality and high cell toxic activity CIK cell. This invention increases augmentation multiple and decreases toxic side effect of single cytokine, which can make cell toxic activity maintain 2-3 weeks.

Description

technical field [0001] The invention relates to a preparation method of cells. Background technique [0002] Cytokine induced killer cells (CIK cells) are a group of heterogeneous cells obtained by culturing mononuclear cells of human peripheral blood, umbilical cord blood or bone marrow with cytokines for a period of time in vitro. Because this kind of cell expresses two kinds of membrane protein molecules, CD3 and CD56, it is also called NK cell-like T lymphocyte, which has both the strong anti-tumor activity of T lymphocytes and the non-major histocompatibility complex of NK cells (major histocompatibility complex, MHC) limited tumor killing advantages. However, existing CIK cells have poor proliferative ability and low cytotoxic activity. Contents of the invention [0003] The object of the present invention is to provide a preparation method of CIK cells with high proliferation and high cytotoxicity in order to solve the problems of poor proliferation and low cytoto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/08C12N5/078
Inventor 王志华秦莉
Owner HARBIN MEDICAL UNIVERSITY
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