CTLL-2细胞的复苏及培养条件的探讨

An Approach into the Condition for Resuscitation and Cultivation of CTLL-2 Cells

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 approach into the optimal condition for resuscitation and cultivation of CTLL 2 cells (cytotoxic T lymphocyte line 2) and to solve the problem of refrigeration by liquid nitrogen.Methods:CTLL 2 cells resuscitated from liquid nitrogen refrigeration were divided into 2 groups,G1 being the experimental group and G2 the control group.The guantity of interleukin 2 in the culture system of G2 was increased to make the terminal concentration 1000 U/ml,and the terminal concentration of IL 2 in that of G2 was made 300 U/ml.The samples were cutured at 37℃,5%CO 2 and observed after 24 hr.Results:Comparing with G2,the G0 phase of CTLL 2 cell in G2 was shortened, the refractive index was intensive; there was proliferation of cells and the morphology of cells was much better than that of G2.One week after resuscitation,G1 entered cell eycle whereas the amount of cells in G2 was markedly reduced,the cells atrophied and finally died.It is worth mention that after entering normal cell cycle,IL 2 contents in the culture system decreased gradually step by step and maintained a constant quantity.The CTLL 2 cells were still proliferating and hereditable steadily.At that time the CTLL 2 cells might be used to detect the reactivity of IL 2.Establishment of this method of cultivation has solved the difficulties in CTLL 2 cell resuscitation in laboratories.