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Preparation method of Tlr7a-induced enhanced DC-NK cells

An enhanced technology of NK cells, applied in the field of biomedicine, can solve the problems of insignificant cytotoxic activity and achieve the effects of low production cost, increased killing activity, and easy control of conditions

Inactive Publication Date: 2016-11-23
SHENZHEN HORNETCORN BIOTECH
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AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the deficiencies of the existing cell technology, and provide a preparation method of Tlr7a (structural formula shown in the figure below) induced enhanced DC-NK cells and its Application, solves the problems of the prior art DC-NK cell culture, the improvement of cytotoxic activity is not significant, etc.

Method used

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  • Preparation method of Tlr7a-induced enhanced DC-NK cells
  • Preparation method of Tlr7a-induced enhanced DC-NK cells
  • Preparation method of Tlr7a-induced enhanced DC-NK cells

Examples

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Embodiment 1

[0027] In Example 1, the obtained PBMCs were isolated and cultured. Include the following steps:

[0028] Collect 100ml of peripheral blood from the patient's venipuncture, transfer it to an anticoagulant tube containing sodium heparin, mix it evenly up and down, and obtain mononuclear cells through density gradient centrifugation of lymph separation fluid. The specific steps are: centrifuge the uniformly mixed whole blood at 1500 rpm for 10 minutes, absorb the upper layer liquid, that is, plasma, inactivate at 56°C for 30 minutes, centrifuge at 2500 rpm for 10 minutes, and set aside. Dilute the precipitated whole blood cells with normal saline 1:1, add the human lymphocyte separation medium and the diluted whole blood cell liquid into the centrifuge tube at a ratio of 1:2, centrifuge at 2000 rpm for 20 minutes, absorb the buffy coat layer, wash with normal saline Wash twice at 1600 rpm and 1300 rpm, respectively, and centrifuge for 7 minutes to obtain mononuclear cells.

[...

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Abstract

The invention provides a preparation method of Tlr7a-induced enhanced DC-NK cells. According to the technical scheme, the DC-NK cells are high-activity lymphocytes obtained by being separated from peripheral blood of a patient, adding a series of cell factors into an in-vitro culture system and conducting Tlr7a stimulation, the DC-NK cells are massively amplified and activated through the culture method, therefore, the tumor-cytotoxicity capacity of the DC-NK cells is improved, and the DC-NK cells reach the level of being applied to clinical tumor treatment. Compared with DC-NK cells prepared through a common method, the DC-NK cells prepared through the method have the advantage that the killing effect is significant and obviously higher than that of the DC-NK cells cultured through the conventional method. According to the method, a large quantity of DC-NK cells which are safe in application and higher in killing activity can be obtained under the in-vitro culture condition, and meanwhile the advantages that the preparation cost is low, the processes are simple, the conditions are easy to control, the requirements on equipment are low, and large-scale production is easy are achieved.

Description

technical field [0001] The invention belongs to the fields of biomedicine, cellular immunology and tumor immunotherapy, and relates to a preparation method of Tlr7a-inducible enhanced DC-NK cells and an application of anti-tumor adoptive immunotherapy. Background technique [0002] Tumor is a major disease that plagues human health, but due to its high complexity, diversity, variability and heterogeneity, an ideal treatment method has not been found. Scientists have invented adoptive immunotherapy for tumor patients who cannot independently produce more effective killing effector cells. Adoptive immunotherapy is a hotspot in the research of tumor treatment, and it has been widely used in clinic. [0003] NK cells or natural killer cells mainly express CD3 - CD56 + Phenotype is an important part of the natural immune system and the body's first line of defense against tumors and infections. Unlike T lymphocytes, NK cells can directly kill tumor cells and virus-infected ce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783C12N5/0784
CPCC12N5/0646C12N5/0639C12N2501/2301C12N2501/2302C12N2501/24C12N2501/50C12N2501/515
Inventor 高东李旺陈艳媛王宇环魏志璋罗晓玲
Owner SHENZHEN HORNETCORN BIOTECH
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