Preparation and use of human body immunocompetent cell DCCIK anti-tumor cell preparation

An anti-tumor cell and immune activity technology, which is applied in the field of preparation of human immune activity cell preparations, can solve the problems of unavailable quantity and systemic toxicity, and achieve the effect of preventing metastasis, recurrence and good clinical application prospects

Inactive Publication Date: 2007-07-04
SHANGHAI ZHONGKE YINGDA BIO TECH
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Problems solved by technology

[0006] (1) Although LAK has achieved great success in mouse tumor experiments, it can produce systemic toxicity due to its severe dependence on IL-2 in clinical applications, which limits its wide application in humans
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Method used

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Examples

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Embodiment Construction

[0041] Below by specific embodiment, the present invention will be further described:

[0042] 1. Preparation of Peripheral Blood Mononuclear Cells (PBMCs)

[0043] Under sterile conditions, anticoagulant peripheral blood from patients with confirmed tumors was taken with a blood separator, diluted with normal saline (0.9% NaCl), and then centrifuged with lymphocyte separation medium (Ficoll, 1.077) (2000rpm × 25'). The PBMCs were isolated, washed twice with Hanks solution, and the number of cells was counted under a microscope. Finally, the cell density was adjusted to 4×10 with serum-free medium (AIM-V). 6 / ml cell suspension.

[0044] 2. Non-adherent vs. Adherent Cell Isolation

[0045] Transfer the cell suspension into a 6-well plate (Nuclon), incubate at 37°C, 5% CO2 for 2-6 hours, then gently flush the cells with a pipette, and collect the non-adherent cell suspension in a centrifuge tube for CIK induction The cells are ready for use; while the adherent cells are left...

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Abstract

The invention discloses the human body immunocompetent cell (DCCIK) anti-tumor cell agent and preparation methods. The method induces patients peripheral blood to DC and CIK using relevant cytokine, mixes the DC and CIK cells according proportion and cultures together after using alpha-Galcer or CI repeatedly impact DC, then gets DCCIK cells. Compared with homologous CIK, its proliferative activity and cytotoxicity are improved. The DCCIK has specific targeting and highly effective wide spectrum kill tumor effect without relevant tumor antigen impact. It can prevent tumour transition and recurrence after tumour surgery, radiotheraphy and chemotherapy. It can extend patient survival time and improve life quality with the assistance of other therapies.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method and application of a human immunocompetent cell (DCCIK) preparation. Background technique: [0002] Immunotherapy has been recognized as the most important means besides tumor surgery, chemotherapy and radiotherapy. It plays an important role in eliminating micrometastasis tumor cells in tumor patients, preventing tumor metastasis and recurrence, and improving patient survival rate and quality of life. Tumor-specific active immunotherapy and passive adoptive immunotherapy research. In the past 20 years, great progress has been made: (1) the identification and purification of cytokines with immune activation function (Miller DL, et al.Science.1982; 215:689-390), which can enhance the body's immunity to cancer cells answer. (2) Develop a method that can selectively and efficiently expand specific immune effector cells, that is, obtain a large number ...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K35/14A61P35/00C12N5/08A61K35/17C12N5/078C12N5/0784
Inventor 刘祥麟王定华
Owner SHANGHAI ZHONGKE YINGDA BIO TECH
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