Culture method for functionally enhanced TILs

Abstract

The invention discloses a culture method for functionally enhanced tumor infiltrating lymphocytes (TILs). The method includes the following steps: separating lymphocytes from tumor tissue, adding a start culture medium, performing inoculation in a 12-well culture plate (4 ml / well), performing start lymphocyte culture for 10 days or 14 days to obtain start TILs, and performing cryopreservation on the obtained start TILs for standby application; suspending the lymphocytes in a 25 cm<2> culture flask (20 ml / flask) by using an induction culture medium, placing the culture flask in an incubator having 5% CO2 at 37 DEG C, and performing TIL induction culture for 1 day; performing half quantity change by using an expansion culture medium, and performing expansion flask culture and expansion bag culture for 13 days or 14 days; on the 14th or 15th day of culture, collecting TILs, performing washing by using normal saline, and resuspending the TILs by using a function enhancement culture medium,and performing incubation for 30 min; and collecting TILs, and performing functional test to obtain the functionally enhanced TILs. The culture method described in the invention can obtain the functionally enhanced TILs with stronger tumor cell killing activity and higher-level anti-tumor cytokine secretion ability.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a cell culture method, in particular to a function-enhanced tumor infiltrating lymphocyte (Tumor infiltrating lymphocytes, TIL) cell culture method. Background technique [0002] Tumor immunotherapy is expected to become a new weapon to cure tumors, especially the application of anti-PD-1 antibody, anti-PD-L1 antibody and anti-CTLA-4 antibody in clinical tumor immunotherapy has received extensive attention, and they significantly prolong the life expectancy of advanced tumors. Survival for patients with malignant melanoma, lung and colon cancers, among others, is showing light for many patients with difficult-to-treat cancers. Immune checkpoint blockers mainly block T cell inhibitory signaling pathways in tumor tissue, thereby continuously activating T cells, breaking through the immune suppression of the tumor microenvironment, and achieving the purpose of killing tumor cells. 2018 is a...

AI Technical Summary

Problems solved by technology

[0005] Due to the limited source of TIL cells, conventional TIL cell culture methods are affected by various factors such as tumor size, the number of TIL cell infiltration in the tumor, and easy contamination during the process of taking tumor tissue, which limits the application of TIL cell therapy in clinical tumor treatment. widely used

Compared with cytokine-induced killer cells (CIK cells) and NK cells, the cell proliferation rate and proliferation rate during TIL cell culture are not as good as the former; and a large amount of IL-2 is required during the culture process, which can promote CD4+CD25+ regulatory Expansion of T lymphocytes, thereby affecting the anti-tumor activity of TIL cells, leading to the occurrence of tumor immune escape

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