Effect of the activation of lymphocyte between freeze-thaw tumor cell and the supernatant of tumorcell culture

Abstract

Objective To compare the effect of freeze-thaw tumor cells and the supernatant from tumor cell culture on the activation of lymphocytes. Methods Malignant melanoma B16 cells were prepared as tumor cell vaccine and the supernatant from tumor cell culture was collected at different time point.Transwell method was used to determine the chemotaxis of lymphocyte attracted by freeze-thaw tumor cells and the supernatant from tumnor cell culture. The cytotoxic activity of lymphocyte was detected by CCK-8 method. Results Freeze-thaw tumor cells and the supernatant from more than 2 h of tumor cell culture were found to show chemotaxis of lymphocyte. The chemotaxis of tumor cell culture more than 4 h was stronger than freeze-thaw tumor cells. Each group of chemotactic lymphocytes demonstrated to have the activity of killing tumor cells. The ability of killing tumor cells induced by the tumor cell culture more than 4 h was stronger than that induced by freeze-thaw tumor cells.In a certain range,the ability of lymphocyte chemotaxis and activation were enhanced over time. Conclusion The chemotaxis and cytotoxic activation of lymphocyte induced by the supernatant from tumor cell culture for a certain time are stronger than those by freeze-thaw tumor cells. The supernatant from tumor cell culture can be used as tumor antigen to get better immune activation instead of the freeze-thaw tumor cells. Key words: Cancer vaccines; Antigens, neoplasm; Tumor cells, cultured; Freeze-thaw tumor cell; Cell culture techniques