动物造模 z-bio 2021-12-13 496

　　Purpose: To obtain data on the phenotype and ratio of NK cells in Bama pig peripheral blood lymphocytes, and to establish a high-efficiency pig CIK (cytokine-induced killer) cell culture method in vitro.

　　Methods: Isolate Bama mini-pig peripheral blood lymphocytes, detect CD2+/CD8+/CD3- cells to characterize the phenotype of pig NK cells; optimize the induced culture conditions, increase the proportion of CIK in induced lymphocytes, and establish high-efficiency CIK cells In vitro induction method.

　　Results: Using optimized induction culture conditions, the proportion of CD3-/CD2+/CD8+ NK cells on the 5th day of induction culture can be as high as 43.63%, which is 5.59 times higher than the proportion of NK cells at the initial isolation; cell proliferation activity experiments show that induction culture Obvious 3 fluorescence peaks can be seen on the 5th day, indicating that the cells have divided 3 times after induction, which is an 8-fold increase in theory compared with the initial separation; the qPCR phenotype analysis of the induced cells shows that the cell population is related to the induction culture on the 5th day. The apparent increase in surface markers is also consistent with the results of flow analysis.

　　Conclusion: An efficient method for inducing and culturing pig CIK cells in vitro has been established.