Study On Cytotoxicity Of Natural Killer Cells Combined With CIK Cells To Breast Cancer Cells

Objective 1.Explore the best method for isolation and culture of natural killer cells(NK cells)from peripheral blood;2.Determine the killing activity of NK cells,CIK cells and NK-CIK combined cells on breast cancer cells and explore the concentration ratio of the two cells with strongest killing activity.Methods 1.Peripheral blood of healthy people was extracted,peripheral blood mononuclear cells(PBMCs)were isolated,NK cells were isolated and cultured by different methods and then selected the culture method with the highest cell purity by flow cytometry.2.Culture cytokine-induced killer cells(CIK cells)using established methods of laboratory.3.The two cells were mixed in different proportions,and the NK cells,CIK cells and the combined cells were used to act on the breast cancer cell lines and the primary breast cancer cells respectively,and the cytotoxic effect of each group was detected by a microplate reader.And find out the cell concentration ratio with the strongest killing activity in the combined cells.Results 1.The cytometry results show that the purity of NK cells obtained by culture after separation of PBMC by microbead is the highest,up to 91.51%,which can be combined as pure NK cells with CIK cells in different proportions.2.The killing rate of NK cells to breast cancer cell lines is 68.23%;the killing rate of CIK cells to breast cancer cell lines is 75.13%.3.When the ratio of NK cells and CIK cells in the combined cells is 1:1,the killing rate of the combined cells on the breast cancer cell lines is the lowest,which is 63.72%,which is lower than the killing rate of the single cells;when the ratio is 1:8,The kill rate is the highest,up to 89.3%,which is higher than the kill rate of single cells.Conclusion 1.Isolation and culture of NK cells and CIK cells in vitro can obtain target cells with high purity,good activity,and a therapeutic amount.2.In vitro,the killing activity of NK cells on breast cancer cells was slightly lower than CIK cells;the combined cells had the highest killing activity when NK:CIK=1:8.