Building method of NK (natural killer) cell and gamma delta T cell co-culture

Abstract

The invention relates to the technical field of cell culture, in particular to a building method of NK (natural killer) cell and gamma delta T cell co-culture capable of realizing the in vitro commonmultiplication culture on NK cells and gamma delta T cells. The method mainly comprises the following steps of S1, reagent selection; S2, performing PBMC culture by a culture medium for 3 days; S3, supplementing the culture medium; maintaining the concentration; performing culture for 4 days; S4, after the seventh day, removing supernatant through centrifugation; transferring the cells into a T25cell culture bottle; then, replacing an EX culture medium; S5, after the tenth day, supplementing the culture medium for maintaining the concentration; S6, after the twelfth day, continuously supplementing the culture medium for maintaining the concentration; S7, after the fourteenth day, detecting the NK cell and gamma delta T cell proportion and the cell total number. The co-culture building method has the advantages that two kinds of anti-tumor immune cells with similar properties are cultured in one step; the cell culture efficiency is improved; the culture cost can be greatly reduced through being compared with that of independent culture of various immune cells for obtaining NK cells and gamma delta T cells; the culture technical difficulty is also reduced.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for establishing co-cultivation of NK cells and γδT cells that can be co-expanded and cultured in vitro. Background technique [0002] Natural killer cells (natural killer cells, NK) are important immune cells in the body, and are closely related to anti-tumor immune technology. First, culture NK cells in vitro and then autologous reinfusion can be used as adjuvant cell therapy for tumor treatment; NK cell culture in vitro is the key to this It is the key link of this kind of cell therapy, and NK cells have intermediate affinity receptors for IL-2, and can undergo a proliferation response under the stimulation of IL-2. The existing NK cells in vitro culture are mainly expanded by adding medium containing IL-2 As a result, there are currently more commercial media to choose from. [0003] γδT cells are T cells that perform innate immune functions. Its TCR is compose...

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Problems solved by technology

[0004] Both γδT cells and NK cells have tumor cell killing activity, and both can be used for adjuvant therapy of tumor patients. It will be of great significance if these two types of cells can be simultaneously expanded and applied to patients. At present, the in vitro expansion and culture of NK cells and γδT cells are carried out separately, and the establishment method of simultaneous expansion and culture is still to be developed, and its significance and value are still to be verified

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