Clinical low-DMSO (Dimethyl Sulfoxide) cryopreservation agent for TCR-T (T-Cell Receptor) cell and cryopreservation method

Abstract

The invention discloses a clinical low-DMSO (Dimethyl Sulfoxide) cryopreservation agent for TCR-T (T-Cell Receptor) cells and a cryopreservation method. The clinical low-DMSO cryopreservation agent for the TCR-T cells comprises the following components: 2.2-5.5%w / v of dimethyl sulfoxide, 0.5-2%w / v of protamine, 0.1-1%w / v of glucan, 0.001-0.01%w / v of vitamin C and 0.5-2%w / v of raffinose, and the volume of the components is fixed to 100ml by using injection water. The method comprises the following steps: 1) directly putting the clinical low-DMSO cryopreservation agent for the TCR-T cells into TCR-T cells, and uniformly mixing so as to obtain a cell suspension; 2) putting the cell suspension into a cryopreservation bag; and 3) putting the cryopreservation bag into a cryopreservation clamp, carrying out programmed freezing to minus 80 DEG C in a velocity of 1 DEG C / minute, and transferring into a refrigerator of minus 80 DEG C for long-term preservation 12-16 hours later.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a low-DMSO cryopreservation agent and a cryopreservation method for clinical-grade TCR-T cells. Background technique [0002] Worldwide, solid tumors are the main cause of death in cancer patients, among which the mortality rates of male patients and female patients accounted for 94.4% and 96.8%, respectively. Traditional treatments such as surgery, radiotherapy and chemotherapy have made great progress in recent years, but this treatment has a great burden on the body, and after malignant metastasis occurs, no matter what traditional treatment is adopted, it is difficult to completely cure of. However, with the rapid development of molecular biology techniques, it has been found that T cells can express T cell receptors (TCRs) or chimeric antigen receptors (CARs) that recognize tumor antigens on the cell surface through gene editing, thereby improving the immune cell funct...

AI Technical Summary

Problems solved by technology

The effect of different cell protectants on the preservation of cells varies greatly

[0012] Traditional TCR-T cell cryopreservation agents contain at least 10% DMSO and macromolecular extracellular protective agents such as dextran. Even excipient-grade DMSO also has side effects such as vascular stimulation and oral mucosal stimulation. , DMSO and macromolecular extracellular protective agents will increase the metabolic burden of the patient's liver, kidney and other organs; at the same time, the effect of cryopreserving TCR-T cells using traditional methods is uneven. It is difficult to maintain cell activity, especially the killing activity against tumor cells is significantly reduced

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