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Method for preparing CIK cell by using heparin anticoagulant plasma

A plasma and cell technology, applied in the biological field, can solve the problems of limited popularization and application, high price of serum-free medium, etc., and achieve the effects of improved biological activity and stable activity.

Inactive Publication Date: 2013-02-13
昆明理工大学附属医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high price of serum-free medium, the clinical application of this method in adoptive immunotherapy is limited.

Method used

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  • Method for preparing CIK cell by using heparin anticoagulant plasma
  • Method for preparing CIK cell by using heparin anticoagulant plasma
  • Method for preparing CIK cell by using heparin anticoagulant plasma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: The method of using the heparin anticoagulated plasma of the patients after cardiac cancer operation to cultivate autologous lymphocytes, the specific content is as follows:

[0031] 1. Clinical data

[0032] Case 1, male, 56 years old, 2 months after surgery for cardiac cancer, was confirmed by pathological examination after surgery.

[0033] 2. Main materials

[0034] Hyclone RPMI-1640 was purchased from Thermo Fisher Biochemical Products (Beijing) Co., Ltd.; recombinant human interleukin-2 (Recombinant human interleukin-2, rhIL-2) for injection was purchased from Beijing Sihuan Biopharmaceutical Co., Ltd.; Recombinant human interleukin-1 alpha (rhIL-1α) was purchased from PeproTech, USA; Recombinant human interferon γ (rhIFN-γ) for injection was purchased from Shanghai Kaimao Biomedical Co., Ltd. Company; anti-human T cell CD3 mouse monoclonal antibody (Monoclonal antibody CD3, CD3McAb) for injection was purchased from Wuhan Institute of Biological Pr...

Embodiment 2

[0042] Embodiment 2: The method of using the heparin anticoagulated plasma of the patient after left breast cancer operation to cultivate autologous lymphocytes, the specific content is as follows:

[0043] 1. Clinical data

[0044] Case 2, female, 54 years old, left breast cancer after 1 year, was confirmed by pathological examination after surgery.

[0045] 2. Main materials

[0046] Hyclone RPMI-1640 was purchased from Thermo Fisher Biochemical Products (Beijing) Co., Ltd.; recombinant human interleukin-2 (Recombinant human interleukin-2, rhIL-2) for injection was purchased from Beijing Sihuan Biopharmaceutical Co., Ltd.; Recombinant human interleukin-1 alpha, rhIL-1α) was purchased from PeproTech, USA; recombinant human interferon γ for injection (Recombinat human interferon γ, rhIFN-γ) was purchased from Shanghai Kaimao Biopharmaceutical Co., Ltd. ; Anti-human T cell CD3 mouse monoclonal antibody (Monoclonal antibody CD3, CD3McAb) for injection was purchased from Wuhan ...

Embodiment 3

[0060] Embodiment 3: The method of using heparin anticoagulated plasma of patients with gastric cancer to cultivate autologous lymphocytes, the specific content is as follows:

[0061] 1. Clinical data

[0062] Case 3, male, 50 years old, gastric cancer patient.

[0063] 2. Main materials

[0064] Hyclone RPMI-1640 was purchased from Thermo Fisher Biochemicals (Beijing) Co., Ltd. Recombinant human interleukin-2 (rhIL-2) for injection was purchased from Beijing Sihuan Biopharmaceutical Co., Ltd. Recombinant human interleukin-1 alpha (rhIL-1 alpha) was purchased from PeproTech, USA. Recombinant human interferon γ for injection (Recombinat human interferon γ, rhIFN-γ) was purchased from Shanghai Kaimao Biopharmaceutical Co., Ltd. Anti-human T cell CD3 mouse monoclonal antibody (Monoclonal antibody CD3, CD3McAb) for injection was purchased from Wuhan Institute of Biological Products, and human lymphocyte separation liquid was purchased from Beijing Soleibao Technology Co., Ltd...

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Abstract

The invention discloses a method for preparing a CIK cell by using heparin anticoagulant plasma. The method uses heparin anticoagulant plasma of a cancerous person to induce and amplify lymphocytes in vitro. The method provided by the invention can expanse the CIK cell greatly. The experiment result shows that the number of CIK cells increases significantly from the seventh day to 21th day, and can be up to 40+ / -15.0 times of the start number of cells. The number of immune phenotype CD3+CD56+ cells in the CIK cells accounts for high proportion in the total number of cell, and can be up to 10-60% when the cells are cultured to the seventh days to 21th days. The biological activity of the CIK cells is improved. The experiment result shows that the cell killing activity is more than or equal to 70% when the cells are cultured to the seventh days to 21th days.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing CIK cells by using blood plasma anticoagulated with heparin. Background technique [0002] Cytokine-induced killer cells (Cytokine-induced killer, CIK) is the human peripheral blood mononuclear cells (Peripheral blood mononuclear cell, PBMC) in vitro by a variety of cytokines (such as anti-CD3McAb, IL-2, IFN-γ, IL -1α, etc.), a group of heterogeneous cells mainly characterized by CD3+CD56+ induced by combined induction, have both the anti-tumor activity of T lymphocytes and the non-major histocompatibility complex (Major Histocompatibility Complex, MHC) restriction of NK cells They are also known as natural killer (NK)-like T lymphocytes (NK-T cells). As a new type of high-efficiency immunocompetent cells, it is considered to be a new generation of anticancer cells because of its incomparable advantages such as strong proliferation ability, broad t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
Inventor 严新民董虹华映坤唐慧高建梅
Owner 昆明理工大学附属医院
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