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J Korean Cancer Assoc > Volume 19(2); 1987 > Article
Journal of the Korean Cancer Association 1987;19(2): 68-79.
MTT 방법을 이용한 림프킨활성 살해세포 ( LAK cell ) 활성도 측정에 관한 연구
박재갑, 권오중, 김선회, 김진복, 함경수, 한문희
Testing of LAK Cell Activity Using a Tetrazolium-based Colorimetric ( MTT ) Assay
Jae Gahb Park, Oh Hoong Kwon, Sun Whe Kim, Jin Pok Kim, Kyung Soo Hahm, Moon H. Han
ABSTRACT
Peripheral blood lymphocytes of normal individuals can be activated by culture with recombinant Interleukin-2 (donated by Genetic Engineering Center, KAIST, Seoul, Korea) leading to expression of cytotoxic activity toward on colorectal carcinoma cell line SNU-C5. After 4 days incubation, the LAK cells were removed by 4 times washing and the number of live target cells in the microtest plates were counted by the tetazolium-based colorimetric (MTT) assay and the survival rate of the target cells was calculated against the control in which the target cells were incubated without Interleukin-2 and effector cells. During 5 days culture, the recombinant KAIST Interleukin-2 induced the potent LAK cells at the concentration of 0.1 U/ml. After 4 days incubation, almost all target cells (SNU-C5) were killed by the LAK cells of 5 days culture when the Effector:Target ratio were 10 : 1 and 5 : l. SNU C5 represented high adhesiveness to the culture flask and microtest plate bottom, so it can be used for the tetazolium based colorimetric (MTT) assay. Tetrazolium-based colorimetric (MTT) assay shows good carrelation between spectrophotometric absorbance and cell number, and it is long-term assay over other cytotoxicity test methods, so semiautomated MTT assay offers a valid. rapid, and simple method to assess cytotoxicity of LAK cells.
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