The invention belongs to the technical field of
cell culture
in vitro, and particularly relates to a preparation method of high-purity, high-multiplication capacity and high-cytotoxin activity CIK (
cytokine induced kill)
cell. The method comprises the following steps: collecting and separating
peripheral blood mononuclear
cell of a patient, eliminating CD4+CD25+
Treg cell by means of Mini MACS (magnetic
active cell sorting) method, and sorting to obtain CD3+, CD4+ and CD8+T cells; and putting the obtained cells into culture solution containing phytohemagglutinin (PHA), so that the PHA concentration in the suspension liquid is 100ng / ml,
hatching for 24h under the culture condition of 5% CO2 at 37 DEG C, transferring the hatched suspension liquid into a
cell culture bottle coated by CD3
monoclonal antibody (1mug / ml), adding IFN (
interferon)-gamma (1000U / ml), adding IL (
interleukin)-2(500U / ml) and IL (
interleukin)-21(1000U / ml) after 48h, compensating
sodium selenite-containing (0.005mg / L)
cell culture after four days, and continuously culturing for 7-14 days to obtain the high-purity, high-multiplication capacity and high-cytotoxin activity CIK (
cytokine induced kill) cell. The quantity, the activity and the purity of the CIK cell which is prepared by the method and amplified
in vitro are improved, so that the antineoplastic function of the CIK is enhanced.