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目的:比较Percoll和Ficoll两种淋巴细胞分离液对肿瘤浸润淋巴细胞纯度及细胞增殖的影响。方法:用Percoll及Ficoll两种淋巴细胞分离液分离H22肝癌小鼠皮下移植瘤内淋巴细胞,对细胞首先行锥虫蓝染色后在显微镜下计算细胞总数和存活率,最后用流式细胞仪分析淋巴细胞纯度及分裂、增殖能力。结果:Percoll及Ficoll分离所得TIL总数分别为(3.28±0.78)×107和(2.02±0.76)×107;Percoll分离所得第3、5、7天的TIL增殖率分别为(22.8±2.31)%、(39.1±1.94)%、(69.85±1.68)%;Ficoll分离所得第3、5、7天的TIL增殖率分别为(13.25±1.68)%、(23.7±5.31)%、(41.5±1.56)%。Percoll分离所得TIL中CD3+T、CD4+T、CD8+T细胞分别为(73.2±6.8)%、(49.1±10.2)%、(22.0±1.4)%;Ficoll分离所得TIL中CD3+T、CD4+T、CD8+T淋巴细胞分别为(47.9±5.0)%、(29.1±8.4)%、(19.7±5.0)%。结论:与Ficoll相比,Percoll分离所得H22肝癌小鼠皮下移植瘤TIL的细胞得率和纯度更高、增殖力较强,将更有利于后续研究。
Objective: To compare the effects of two Percoll and Ficoll lymphocytes on the purity of tumor-infiltrating lymphocytes and cell proliferation. METHODS: Peripheral blood lymphocytes were subcutaneously transplanted from H22 hepatocarcinoma mice using Percoll and Ficoll. The cells were stained with trypan blue first to calculate the total number of cells and the survival rate under microscope. Finally, the expression of T lymphocytes was analyzed by flow cytometry Lymphocyte purity and division, proliferation. Results: The total number of TILs isolated by Percoll and Ficoll was (3.28 ± 0.78) × 107 and (2.02 ± 0.76) × 107, respectively. The proliferation rates of TIL on the 3rd, 5th and 7th day after Percoll isolation were (22.8 ± 2.31)%, (39.1 ± 1.94)% and (69.85 ± 1.68)%, respectively. The proliferation rates of TIL on the 3rd, 5th and 7th days after Ficoll separation were (13.25 ± 1.68)%, (23.7 ± 5.31)% and (41.5 ± 1.56) . (73.2 ± 6.8)%, (49.1 ± 10.2)% and (22.0 ± 1.4)%, respectively, in TIL obtained from Percoll. CD3 + T, CD4 + T and CD8 + T lymphocytes were (47.9 ± 5.0)%, (29.1 ± 8.4)% and (19.7 ± 5.0)%, respectively. CONCLUSION: Compared with Ficoll, the TIL obtained from Percoll isolated H22 hepatocarcinoma mice has higher cell yield and purity, stronger proliferative capacity, and will be more conducive to follow-up studies.