THE PROTOCOL FOR ANALYZING THE POTENCY OF POST-EXPANDED HUMAN NK CELLS IN KILLING CANCER CELL LINE IN VITRO

Trọng Phúc Nguyễn1,, Thế Hải Phùng1, Hoàng Phương Nguyễn1, Ngọc Tuấn Nguyễn1, Trung Kiên Hoàng1, Thu Hằng Ngô1, Văn Mão Cấn1, Lĩnh Toàn Nguyễn1, Văn Đông Lê1, Khắc Đại Đỗ1, Anh Tuấn Đỗ2
1 Vietnam Military Medical Academy
2 K Central Hospital

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Abstract

Objective: This study aims to establish a procedure to evaluate the cytotoxicity of NK cells after being activated and proliferated in vitro based on the evaluation tool of flow cytometry system. Subjects and research methods: We analyzed two sources of NK cells: (1) Peripheral blood NK cells isolated from a donor (the patient was diagnosed with prostate cancer at the central K hospital - Tan Trieu campus) at the time before proliferative culture (first day - D0; NK-D0) and (2) NK cells at the time of post-proliferative culture with the NK cell activation culture kit (KBM kit) in 14 days (Day 14 - D14; NK-D14); these two sources of NK cells were co-cultured with prostate cancer cells (PC3) in a ratio of 5:1 (NK:PC3) within 6 hours to evaluate the potency of two NK cells. The percentage of survived PC3 cells that were not lysed by NK cells was assessed by flow cytometry. Results: Based on the established procedure, we found that proliferative-cultured NK cells exhibited a stronger ability to kill cancer cell line (PC3) than fresh NK cells isolated from the patient. Conclusion: We have successfully established a procedure to evaluate the cancer cell killing ability of NK cells after proliferation and activation. This procedure is a premise for clinical application studies to evaluate the potency of immune cells after culture.

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References

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