The invention discloses an NKT
cell culture optimization method, and belongs to the technical field of
cell culture.The method comprises the steps that firstly, PBMC is sorted through a spiral micro-fluidic
chip integrated with CD3 / CD56 double-labeled magnetic beads, and CD3 + CD56 + cells with the purity larger than or equal to 95% are obtained through the synergistic effect of
dielectrophoretic force and a
magnetic field; then, in a GIBCO culture solution added with the IL-2 / IL-15 / alpha-GalCer sustained release microspheres, the pH, dissolved
oxygen and
lactic acid concentration are accurately regulated and controlled through a real-time
monitoring system, and temperature-
oxygen partial pressure gradient culture is adopted; and finally, three times of
centrifugation are combined with NKT4
solution treatment, and the cells are collected in cooperation with a gradient cooling procedure, so that the
cell activity maintenance rate is greater than or equal to 95%. Compared with the traditional technology, the method has the advantages that the NKT
cell sorting purity and activity are remarkably improved, the
culture environment is regulated and optimized through dynamic
metabolism, and accurate control over the culture process is achieved.