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肿瘤浸润淋巴细胞（ＴＩＬ）的增殖力、活力、杀伤力是ＴＩＬ临床治疗的基本问题。由于Ｒｏｓｅｎｂｅｒｇ方法存在着许多不足之处。本实验室对其一些重要步骤进行了改进：①用单一冷胶原酶消化肿瘤组织块；②省去影响ＴＩＬ活力的离心操作；③ＴＩＬ培养４８ｈ内除去掺杂在ＴＩＬ中的肿瘤细胞等。由于除去了影响ＴＩＬ活力的抑制因素，ＴＩＬ的增殖率、生存率、活力、杀伤力明显增加。有８０％肿瘤组织的ＴＩＬ经扩增可超过１×１０￣９细胞数，生长周期曲线反映了冷酶法的增殖率超过Ｒｏｓｅｎｂｅｒｇ方法。本方法的建立，为ＴＩＬ克隆、建株打下了基础，显示了ＴＩＬ临床治疗潜能的提高。
Tumor infiltrating lymphocytes (TIL) proliferation, vitality, lethality is the basic problem of clinical treatment of TIL. Due to the Rosenberg method there are many shortcomings. In our laboratory, some important steps were improved: ①Using a single cold collagenase to digest the tumor tissue; ②Extracting the centrifugation to affect the activity of TIL; ③TEL cells cultured in TIL for 48h were removed. TIL proliferation, survival, vitality, and lethality were significantly increased due to the removal of the inhibitory factors that affect the viability of TIL. TIL of 80% of the tumor tissue can be expanded to more than 1 × 10 ~ 9 cells, the growth cycle curve reflects the cold enzyme proliferation rate over the Rosenberg method. The establishment of this method laid the foundation for the cloning and construction of TIL, which shows the improvement of the clinical therapeutic potential of TIL.