A kind of preparation method of cytotoxic T cell

A cytotoxic and cellular technology, applied in the field of preparation of cytotoxic T cells, can solve the problems of low yield and low purity

Active Publication Date: 2021-03-30
蓝莲(杭州)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The existing method for preparing CTL cells has low yield and low purity

Method used

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  • A kind of preparation method of cytotoxic T cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] CTL cell culture

[0045] 1. Verify that the suction tube of the plasma bag is sealed. Wipe the pipette and scissors with two different cleanroom wipes soaked in 70% ethanol.

[0046] 2. Hold the suction tube with tweezers, cut the front end of the tube and control the flow rate with tweezers to transfer the plasma into a 50ml test tube. Record the plasma volume transferred.

[0047] 3. Centrifuge the tube (400g, 4°C) for 30 minutes to remove platelets.

[0048] 4. Pour the supernatant liquid into a new 50ml test tube.

[0049] 5. Put the test tube into a constant temperature water tank (56°C) and incubate for 30 minutes.

[0050] 6. Centrifuge tubes (500g, 4°C) for 60 minutes to remove precipitated fibrin.

[0051] 7. Pour the upper liquid (autologous serum) into a new 50ml test tube. Label the date of preparation and refrigerate (4°C).

[0052] 8. Take a new plasma bag to confirm that the seal is intact, and shake the plasma evenly.

[0053] 9. Wipe the suctio...

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Abstract

The invention relates to a preparation method of cytotoxic T cells, and belongs to the technical field of cell preparation methods. The preparation method comprises the following steps: 1) conductingPBMC cell culture for plasma; 2) conducting CTL cell culture on the first day; 3) harvesting and freezing the CTL cells. In the preparation method of the cytotoxic T cells, a magnetic bead separationmethod is used to screen and remove unnecessary cells and cooperate with various interleukin and Lymacin-T methods, so that the preparation method of the invention has the advantages of greatly increasing yield and purity of the CTL cells compared with the existing culture methods.

Description

technical field [0001] The invention belongs to the technical field of cell preparation methods, in particular to a preparation method of cytotoxic T cells. Background technique [0002] Cytotoxic T lymphocytes (cytotoxic lymphocytes) specialize in secreting various cytokines to participate in immunity, and have a killing effect on antigenic substances such as certain viruses and tumor cells. It can continuously kill target cells with high efficiency. The existing method for preparing CTL cells has low yield and low purity. Contents of the invention [0003] In view of the above-mentioned problems existing in the prior art, the purpose of the present invention is to design and provide a technical scheme for the preparation method of cytotoxic T cells, which uses magnetic bead separation to screen out useless cells and cooperates with various interleukins and Lymactin- Compared with the existing culture method, the method of T has the advantage of greatly increasing the y...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0638C12N2500/80C12N2501/2302C12N2501/2306C12N2501/2312C12N2501/515
Inventor 杨罕闻
Owner 蓝莲(杭州)生物科技有限公司
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