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Cell separation culture solution and T cell separation culture method

A technology of separation culture and culture medium, which is applied in the field of cell separation culture medium and T cell separation culture, which can solve the problems of detecting T cell performance that affect T cell performance, difficulty in observing T cell growth, and affecting T cell culture environment, etc.

Inactive Publication Date: 2020-05-01
广州航华生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If it is cultured directly, it is not only difficult to observe the growth of T cells during the culture cycle, but also affects the culture environment of T cells, and affects the subsequent detection of T cell performance and T cell performance.

Method used

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  • Cell separation culture solution and T cell separation culture method
  • Cell separation culture solution and T cell separation culture method
  • Cell separation culture solution and T cell separation culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] (1) Isolation of mononuclear cells from whole blood:

[0053] 5 ml of human peripheral venous blood was collected, centrifuged at 2500 rpm for 10 min, and the upper layer of autologous plasma was taken for retention, and then stored in a 4 °C refrigerator after being bathed at 56 °C for 30 min. The volume ratio of the blood cell suspension in the lower layer is 1:1, and 3 ml of normal saline is added to dilute, and the mixture is mixed to obtain a blood cell dilution. Take a new centrifuge tube, add 5ml of neutrophil separation solution according to the volume ratio of blood cell dilution solution of 1:1, slowly add blood cell suspension to the upper layer of neutrophil separation solution, and centrifuge at 550g for 30min. After centrifugation, the layers were separated, the mononuclear cell layer was sucked, and the mononuclear cells were collected by centrifugation at 250 g for 10 min.

[0054] (2) Lysis of red blood cells: Add 5 ml of red blood cell lysis solution ...

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Abstract

The invention relates to the technical field of cell culture, and in particular to a cell separation culture solution and a T cell separation culture method. The cell separation culture solution includes a culture solution A, a culture solution B, and a culture solution C, and the three culture solutions cooperate in synergy and work together, and are conducive to stimulating transformation of monocytes to CIK cells. According to the cell separation culture solution and the T cell separation culture method, by adding neutrophil separation fluid to blood cells, content of neutrophils in the cells is reduced; after mononuclear cells are extracted by using the neutrophil separation liquid, most of red blood cells in mononuclear cell layer are removed by using a red blood cell lysis liquid, content of the red blood cells in the mononuclear cells is obviously reduced, and number of lymphocytes is increased; after the final culture, effective cell proportion in the cells is increased, and tumoricidal activity of cell population is increased.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a cell separation culture medium and a method for T cell separation and culture. Background technique [0002] T cells are derived from lymphoid stem cells derived from bone marrow. After differentiated and matured in the thymus, they are distributed to immune organs and tissues throughout the body through lymphatic and blood circulation to exert immune function. [0003] Adoptive immune cell therapy refers to the isolation of immune active cells from the human body, expansion and functional identification in vitro, and then infusion into the human body, so as to directly kill tumors or stimulate the body's immune response to kill tumor cells. Adoptive T cell therapy mainly includes TIL, LAK, CIK, TCR-T, CAR-T and other categories. [0004] At present, culturing T cells in vitro usually includes the following four steps: isolation of mononuclear cells, activation induction,...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N2501/2301C12N2501/2302C12N2501/24C12N2501/515C12N2509/00
Inventor 马玉国
Owner 广州航华生物医药科技有限公司
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