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目的:建立苏木素染色测定免疫效应细胞介导的细胞毒效应的方法:。方法:采用苏木素染色法观察人贴壁肿瘤细胞系Hep2的增殖活性,测定了人NK细胞系NK92、NKL和YT对SGC7901、Hep2细胞的细胞毒效应,新西兰白兔CTL细胞对VX2细胞的细胞毒效应。同时与MTT比色法及CCK-8比色法进行比较。结果:NK92、NKL和YT三种效应细胞对SGC7901和Hep2细胞杀伤6h或16h后,在效靶比分别为10:1、5:1、2.5:1和1.25:1时,三种方法:所测结果:均具有良好的效靶关系,且杀伤趋势基本一致。6次苏木素染色法测定NK92对SGC7901和Hep2细胞的细胞毒效应结果:显示CV<8%,MTT比色法测定结果:CV<12%,前者具有更好的重复性。苏木素染色法和MTT比色法所测兔CTL杀伤效应结果:一致,线性关系明显。结论:苏木素染色法用于免疫效应细胞对贴壁肿瘤细胞系的细胞毒效应测定,具有特异、稳定、简便、经济等优点。
Objective: To establish a method of hematoxylin staining for measuring the cytotoxicity mediated by immune effector cells. Methods: Hematoxylin-eosin staining was used to observe the proliferation activity of human adherent tumor cell line Hep2. The cytotoxic effects of human NK cell line NK92, NKL and YT on SGC7901 and Hep2 cells were measured. The cytotoxicity of CTL cells in New Zealand white rabbits to VX2 cells effect. MTT colorimetric method and CCK-8 colorimetric method were compared. Results: After killing NK cells by SGC7901 and Hep2 cells for 6h or 16h, NK92, NKL and YT cells were treated with 10: 1, 5: 1, 2.5: 1 and 1.25: Test results: all have good target effect, and the killer trend is basically the same. Cytotoxic effect of NK92 on SGC7901 and Hep2 cells was detected by hematoxylin-eosin staining for 6 times. The results showed that CV <8%. MTT assay showed that CV <12%. The former had better repeatability. Hematoxylin and MTT assay rabbit CTL killing effect results: consistent, significant linear relationship. Conclusion: The hematoxylin staining method is used to determine the cytotoxic effect of immune effector cells on adherent tumor cell lines, which is specific, stable, simple and economical.