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Clinical N-CIK cell culture and quality control and identification kit and application

A kind of N-CIK, kit technology, applied in the fields of immunology, biotechnology and biology, can solve the problems of complex culture method, long cycle, instability and so on

Inactive Publication Date: 2010-12-15
山东迪博生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the existing technology has tried to develop CIK technology in different aspects. For example, CN101506356 increased the ratio of CD3+CD56+ on the basis of LAK technology, and the results disclosed in CN101519646A showed that both CD3+CD56+ and CD8+ T cells were increased. , but the active ingredients of the cells are not comprehensive enough, and there are no corresponding regulatory requirements for the quality control and identification of immune cells, and there is no induction culture in the form of kits, preparation of N-CIK cells, and product quality control identification; but the actual clinical application However, due to the limitations of many reagents and factors used in the preparation, long cycle time and inconsistent standards, the culture process is cumbersome and unstable, and the cell quality is difficult to control and even polluted.
For the preparation of dendritic cell vaccines in the clinical research stage, a simple kit preparation method has been disclosed in CN16091095A, which is expected to improve the problem of complicated culture methods in its clinical application in the future

Method used

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  • Clinical N-CIK cell culture and quality control and identification kit and application
  • Clinical N-CIK cell culture and quality control and identification kit and application
  • Clinical N-CIK cell culture and quality control and identification kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1: Preparation of Human Peripheral Blood Mononuclear Cell Isolation Kit

[0075] One disposable medical 200ml blood collection bag (with special needle for blood collection) purchased from the market, appropriate amount of medical 0.9% normal saline and lymph separation fluid, together with laboratory equipment, disposable Consumables, such as pipettes or dispensers, 50mL centrifuge tubes, 25mL pipettes, and 300mL sealed clean mixing bottles, are cleaned, disinfected or sterilized, and brought into the Class 100 clean room required by GMP for use.

[0076] Its preparation steps are as follows:

[0077] ① Transfer 180mL-200mL of medical 0.9% Shengli saline into a 300mL sealed and clean mixing bottle, tighten the cap and seal it;

[0078] ② Transfer the lymphatic separation solution to a clean 50mL centrifuge tube, add 10mL-15mL near the bottom of each tube, screw the cap tightly and seal it;

[0079] ③In addition, take one disposable blood collection bag (with ...

Embodiment 2

[0081] Example 2: Preparation of Human Peripheral Blood Mononuclear and N-CIK Cell Fluorescence Identification Kit

[0082] Purchase reagents according to Table 1: fluorescent antibodies (FITC-IgG\PE-IgG, FITC-CD3\PE-CD4, FITC-CD14\PE-CD19, FITC-CD3\PE-CD56, FITC-CD3\PE-CD8), Bovine serum (FCS), medical 0.9% physiological salt (NS), phosphate buffer solution (PBS) and paraformaldehyde (PFA), and prepare relevant liquid transfer equipment and disposable consumables in a clean room; complete the preparation by the following methods:

preparation example 1

[0083] Preparation example 1: Peripheral blood mononuclear cell fluorescence identification kit, including 20-30 sets of P-1-3 special fluorescent antibody tubes (brown light-proof) and 1 bottle of washing solution and fixative solution, of which P-1-3 The proportions and volumes of each antibody tube filled with the corresponding fluorescent antibodies are shown in Table 1.

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Abstract

The invention provides a kit system for induction and culture of clinical N-CIK cells and control and identification of product quality and application thereof. The kit system comprises a peripheral blood mononuclear cell separation and identification kit, a clinical N-CIK cell culture, induction and amplification kit and a product quality control and identification kit. The invention provides preparation of the clinical N-CIK cells and a design and a method for the quality control related kit system. According to the kit system, the clinical application of the N-CIK cells is more stable, simpler and more efficient. The invention provides a stable method general for clinical T cell preparations for production, quality control and identification in a kit mode. According to the method provided by the invention, NK and CIK cells cultured by the kit are applied to clinical comprehensive tumor treatment so as to clear residual focus, improve the treatment effect and prevent relapse and transfer of tumor.

Description

technical field [0001] The invention relates to the fields of biotechnology and biology and immunology. In particular, the present invention relates to a kit for the induction, cultivation and product quality control identification of NK and CIK (ie N-CIK) cells and the application of N-CIK cells in comprehensive treatment of tumors. Background technique [0002] Tumor, as one of the important causes of human death, has become the primary problem threatening human health and has received widespread attention from society. With the continuous development of tumor treatment methods, the comprehensive treatment of surgery, radiotherapy, chemotherapy and combined biological therapy has become a new perspective of tumor treatment. [0003] Although the existing tumor treatment methods still cannot completely cure the tumor, the use of biological immunology technology to carry out tumor immunotherapy to improve the quality of life and survival of tumor patients has been increasin...

Claims

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Application Information

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IPC IPC(8): C12N5/078C12N5/0783G01N33/53G01N21/64C12Q1/02A61K35/14A61P35/00A61K35/15
Inventor 徐峰波
Owner 山东迪博生物科技股份有限公司
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