论文部分内容阅读
目的:探讨CTLL-2细胞(细胞毒性T淋巴细胞株)的复苏及培养的最佳条件,解决实验室CTLL-2细胞经液氮冻存后复苏难的问题。方法:将从液氮取出复苏后的CTLL-2细胞分为两组,在实验组培养体系中加大IL-2的剂量,使终浓度为1000U/ml;对照组培养体系中IL-2剂量终浓度为300U/ml,培养观察。结果:实验组与对照组比较实验组CTLL-2细胞静止期缩短,细胞折光率强,有增殖现象,细胞形态明显优于对照组。实验组复苏一周后进入正常细胞周期,而对照组细胞数量减少,细胞萎缩最终死亡。值得一提的是实验组进入正常细胞周期后逐渐递减培养体系中的IL-2含量,最后维持在一个恒定的剂量上,CTLL-2细胞仍然稳定增殖传代,这时的CTLL-2细胞可以用来检测IL-2的反应性,这种培养方法的建立解决了实验室CTLL-2细胞复苏难的问题。
Objective: To investigate the best conditions of CTLL-2 cells (cytotoxic T lymphocytes) in resuscitation and culture, and to solve the problem of difficult recovery of CTLL-2 cells after cryopreservation in laboratory. Methods: The CTLL-2 cells from the recovery of liquid nitrogen were divided into two groups, the dose of IL-2 was increased in the experimental group culture system to make the final concentration of 1000U / ml; the IL-2 dose The final concentration of 300U / ml, cultured observation. Results: Compared with the control group, CTLL-2 cells in experimental group were shortened in quiescence, with strong refractive index and proliferation, and the cell morphology was significantly better than the control group. The experimental group entered the normal cell cycle one week after the resuscitation, while the number of the control group decreased and the cell atrophy eventually died. It is worth mentioning that the experimental group after entering the normal cell cycle gradually decreased culture system IL-2 content, and finally maintained at a constant dose, CTLL-2 cells are still stable proliferation of passage, when the CTLL-2 cells can be used To detect the reactivity of IL-2, this culture method to establish the laboratory CTLL-2 cells to solve the problem of difficult recovery.