A method for in vitro expansion of anti-tumor T cells derived from non-sentinel lymph node
A sentinel lymph node and lymphocyte technology, applied in the medical and health field, can solve the problems of reducing the anti-tumor ability of T cells, polarization of T cell subsets, weak anti-tumor function, etc. , the effect of maintaining anti-tumor activity
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[0055] The number, proliferation efficiency, expression of immunosuppressive molecules and immune activation molecules and anti-tumor ability of non-sentinel lymph node-derived anti-tumor T cells amplified by Example 3 of the present invention were compared with those amplified by conventional methods, specifically: :
[0056] 1. About the number of cells:
[0057] Take the quantity as 1.6*10 6 Anti-tumor T cells derived from non-sentinel lymph nodes were expanded in vitro by the method described in Example 3 of the present invention, counted after the culture was completed, and the total amount of anti-tumor T cells derived from non-sentinel lymph nodes was 413*10 6 indivual.
[0058] Take the same quantity as 1.6*10 6 The non-sentinel lymph node-derived T cells were expanded in vitro by conventional methods, counted after the culture was completed, and the total amount of non-sentinel lymph node-derived anti-tumor T cells obtained was 45.6*10 6 indivual.
[0059] The re...
Embodiment 1
[0074] A method for in vitro expansion of anti-tumor T cells derived from non-sentinel lymph nodes, comprising the following steps:
[0075] (1) Antigen sensitization and immunosuppressive blockade:
[0076] a, get the lymphocyte culture medium that is 100:2 in volume ratio, fetal bovine serum mixes, obtain lymphocyte culture medium, fetal bovine serum mixture; Add interleukin 2 in lymphocyte culture medium, fetal bovine serum mixture, adjust all The concentration of said interleukin 2 in lymphocyte culture medium and fetal bovine serum mixture is 100 units / ml, to obtain culture medium A, for subsequent use;
[0077] b. Take part of the culture medium A, add non-sentinel lymph node-derived anti-tumor T cells, adjust the volume of the culture medium, so that the density of the non-sentinel lymph node-derived anti-tumor T cells in culture medium A is 1*10 6 cells / ml, cultivated for 12 hours;
[0078] c. Another part of the culture solution A was taken, and anti-programmed deat...
Embodiment 2
[0100] A method for in vitro expansion of anti-tumor T cells derived from non-sentinel lymph nodes, comprising the following steps:
[0101] (1) Antigen sensitization and immunosuppressive blockade:
[0102] a, get the lymphocyte culture medium that is 100:5 in volume ratio, fetal bovine serum mixes, obtain lymphocyte culture medium, fetal bovine serum mixture; Add interleukin 2 in lymphocyte culture medium, fetal bovine serum mixture, adjust all The concentration of interleukin 2 in lymphocyte culture medium and fetal calf serum mixture is 1000 units / milliliter to obtain culture medium A for subsequent use;
[0103] b. Take part of the culture medium A, add non-sentinel lymph node-derived anti-tumor T cells, adjust the volume of the culture medium, so that the density of the non-sentinel lymph node-derived anti-tumor T cells in culture medium A is 2*10 6 cells / ml, cultivated for 24 hours;
[0104] c. Another part of the culture solution A was taken, and anti-cytotoxic T lym...
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