Real time visualization of cancer cell death, survival and proliferation using fluorochrome-transfected cells in an IncuCyte® imaging system
Real time visualization of cancer cell death, survival and proliferation using fluorochrome-transfected cells in an IncuCyte® imaging system
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DOI：
10.14440/jbm.2020.323
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发表时间：
2020-01-01
期刊：
影响因子：
--
通讯作者：
Ruth, Jeffrey H.
中科院分区：
文献类型：
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作者：
Lanigan, Thomas M.;Rasmussen, Stephanie M.;Ruth, Jeffrey H.
癌症免疫疗法与更传统的治疗形式一样，是一种快速发展且可行的癌症治疗方法。检查癌细胞与免疫系统细胞之间动态相互作用的实时细胞分析技术对于评估新疗法变得越来越重要。在本报告中，我们使用IncuCyte (R)成像系统来研究各种免疫细胞对癌细胞系的杀伤潜力。 IncuCyte (R) 系统跟踪由红色荧光蛋白标记的活细胞和细胞死亡，如 caspase-3/7 试剂所示，该试剂在凋亡途径激活时会产生绿色荧光信号。尽管这种方法很有效，但获得商业荧光癌细胞系的成本很高，而且可用的细胞系范围有限。为了克服这一障碍，我们开发了一种廉价的方法，使用表达核定位 mKate2 红色荧光蛋白的慢病毒构建体来稳定地标记癌细胞。我们证明这种方法可以有效地标记多种细胞系，从而可以分析不同的癌症以及同一类型癌症的不同细胞系。
Cancer immunotherapy is a rapidly advancing and viable approach to treating cancer along with more traditional forms of therapy. Real-time cell analysis technologies that examine the dynamic interactions between cancer cells and the cells of the immune system are becoming more important for assessment of novel therapeutics. In this report, we use the IncuCyte (R) imaging system to study the killing potential of various immune cells on cancer cell lines. The IncuCyte (R) system tracks living cells, labeled by a red fluorescent protein, and cell death, as indicated by the caspase-3/7 reagent, which generates a green fluorescent signal upon activation of apoptotic pathways. Despite the power of this approach, obtaining commercially fluorescent cancer cell lines is expensive and limited in the range of cell lines that are available. To overcome this barrier, we developed an inexpensive method using a lentiviral construct expressing nuclear localized mKate2 red fluorescent protein to stably label cancer cells. We demonstrate that this method is effective in labeling a wide variety of cell lines, allowing for analyses of different cancers as well as different cell lines of the same type of cancer.