Tumor infiltrating lymphocytes separation method

Abstract

The invention relates to a cells separating technology, and provides a tumor infiltrating lymphocytes separation method. The method is characterized by adding tumor tissue masses in a trypsin-EDTA solution for immersion and incubation, fully digesting and dispersing the tumor tissue masses; separating and collecting single cell and being resuspended by the trypsin-EDTA solution for multitime density gradient centrifugation, incubating and separating by an erythrocyte lysate, using calf serum-containing PBS buffer or resuspending to obtain the dissolved tumor infiltrating lymphocytes. In the invention, an enzymatic digestion method and a machinery method are combined for dual separating the enzymatic digestion with high efficiency, compared with the machinery method or the enzymatic digestion method, the survival rate of the obtained tumor infiltrating lymphocytes is high, and the method is simple and easy to operate. The obtained tumor infiltrating lymphocytes can be massively propagated through in vitro stimulation of interleukin 2 (1L-2), have antineoplastic activity with specialty and high efficiency, and provides powerful basis and new approach for treating tumour.

Description

technical field [0001] The invention belongs to cell separation technology, and relates to a method for separating tumor infiltrating lymphocytes. Background technique [0002] In 1986, Rosenberg et al. first reported lymphocytes isolated from mouse solid tumors, called "tumor infiltrating lymphocytes (TIL)", which are mainly T cells that mainly exist in the tumor stroma. A heterogeneous population of lymphocytes, mostly clustered around the tumor or in the stroma surrounding the cancer nest. Further studies have found that this kind of cells can proliferate in large quantities after being stimulated by interleukin 2 (IL-2) in vitro, and has a strong effect of specifically killing tumor cells, and its anti-tumor effect is better than that of lymphokine-activated killer cells ( Lymphokine activated killer cell, LAK) is 50-100 times higher. For example, studies have found that TILs from melanoma patients only have killing activity on autologous tumor cells, but not on autolo...

AI Technical Summary

Problems solved by technology

[0004] At present, most TILs are separated by mechanical methods or enzymatic digestion methods, which not only cost a lot of manpower and material resources, but also reduce the cell activity of TILs after separation.