Optimization Of The Method To Cultivate NK Cells From Abandoned White Cells

Abscract Objective:Using abandoned white cells separated from preparation of blood production to cultivation NK cells in vitro, and testing the function of the NK cells, optimize the method of cultivation allogeneic NK cell for clinical application.Methods:Abandoned white cells separated from blood production were collected from Shanxi Blood Center during March to November 2014, 15 cases of healthy donors; PBMCs were isolated from the abandoned white cells and cultured for 17 days using culture bottles as previously coated Antibody(group A was coated with CD3 m Ab, group B was coated with CD16 m Ab and group C was coated with CD3 m Ab and CD16 m Ab); immunophenotypic analysis and cell surface receptors of the PBMCs and expanded NK cells was conducted using flow cytometry; the anti-tumor cytoxicity against K562 and Raji cells was measured using LDH cytotoxicity assay and flow cytometry.Results:After expansion for 17 days,compared with preamplification, the proportions of NK cells, group A[(15.19±12.22)% vs(16.34±10.51)%,p<0.05], group B[(83.63±10.63)%vs(16.34±10.51)%,p<0.05], group C[(49.40±12.64)% vs(16.34±10.51)%,p<0.05]; number of NK cells were more than 109. The expression ratio of NK cell surface activated receptor NKp30 and NKp46 were significantly increased,NKG2D’s rate changes were not statistically significant. The expansion NK cells had great cytotoxicity activity against K562 cells.Conclusion:Abandoned white cells from preparation of blood production after cultivated for 17 days, NK cells’ rate could reach 90%, cell number were more than 109, NK cells’ activity were reinforced, and cytotoxicity activity against tumor cells were increase, which could reach the standard clinical application.