A kind of culture medium and activation method of T lymphocyte

A lymphocyte and activation method technology, applied in the field of biological cell culture, can solve the problems of low purity and low efficiency, and achieve the effect of improving efficiency and purity, and increasing the number of cell proliferation

Active Publication Date: 2022-05-31
蓝莲(杭州)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing preparation of activated T lymphocytes generally has problems such as low efficiency and low purity

Method used

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  • A kind of culture medium and activation method of T lymphocyte

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Experimental program
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Embodiment 1

[0037] 1. Confirm that the suction tube of the plasma bag is sealed. Use two different cleanroom wipes soaked in 70% ethanol

[0039] 3. Centrifuge the tube (400 g, 4°C) for 30 minutes to remove platelets.

[0040] 4. Pour the upper liquid into a new 50ml test tube.

[0041] 5. Put the test tube into a constant temperature water tank (56° C.) and incubate for 30 minutes.

[0042] 6. Centrifuge the tube (400 g, 4°C) for 60 minutes to remove precipitated fibrin.

[0043] 7. Pour the supernatant (autologous serum) into a new 50ml tube. Label the patient's name on the test tube, prepare

[0044] 8. Confirm that the suction tube of the PBMC bag is sealed. Hang the PBMC bag. Use two different inoculations in 70% ethanol without

[0045] 9. Transfer cells into 50 ml tubes. Record the volume of the transfer.

[0046] 10. Calculate the number of 50ml test tubes required, and add 15ml of lymphocyte separation liquid 1077 to all the test tubes.

[0047] 11. Prepare enough new 50ml tubes t...

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Abstract

The invention discloses a culture solution and an activation method for T lymphocytes. The culture solution includes: PBMCs culture solution and AT culture solution; the PBMCs culture solution includes: 10ml-20ml ALyS505N-0; the AT culture solution includes: 10-15ml 5ug / ml Lymactin‑T anti-CD3 monoclonal antibody, 4‑6ml of ALyS505N‑0, 60‑80ul of 5x10 5 IU / ml IL‑2, 10‑20ul of 5x10 5 IU / ml IL-6, 300-450mg isoflavones, 10-15mg vitamin B6, 3-5ml autologous serum; the present invention found isoflavones, vitamin B6 and Lymactin-T anti-CD3 monoclonal antibody, IL-2, IL-6 The cooperation can promote the rapid proliferation of T lymphocytes in the induction period, and improve the efficiency of activating T lymphocytes, so that the preparation efficiency and purity of activated T lymphocytes are high.

Description

A kind of culture medium and activation method of T lymphocyte technical field The present invention relates to biological cell culture field, particularly a kind of culture liquid and activation method of T lymphocyte. Background technique [0002] T lymphocytes are derived from pluripotent stem cells of the bone marrow (from the yolk sac and liver in the embryonic stage). in human embryos During the early and primary stages, some pluripotent stem cells or pre-T cells in the bone marrow migrate into the thymus, and are differentiated under the induction of thymic hormones. mature into immunocompetent T cells. Thymus-dependent distribution of mature T cells to peripheral immune organs through the bloodstream It can be recirculated through lymphatic vessels, peripheral blood and tissue fluid to play the functions of cellular immunity and immune regulation. T The recycling of cells is conducive to extensive contact with antigenic substances entering the body, strength...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N2501/999C12N2501/515C12N2501/2302C12N2501/2306C12N2500/38C12N2500/32
Inventor 杨罕闻
Owner 蓝莲(杭州)生物科技有限公司
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