In this study we describe three different methods for labeling T lymphocytes with cell trace violet (CTV), in order to track cell division in mouse and human cells, in both the in vitro and in vivo setting. We identified a modified method of CTV labeling that can be applied directly to either conventional or spectral flow cytometry, that maintained lymphocyte viability and function, yet minimized dye spill‐over into other fluorochrome channels. Our optimized method for CTV labeling allowed us to identify up to eight cell divisions and the replication index for in vitro‐stimulated mouse and human lymphocytes, and the co‐expression of T‐cell subset markers. Furthermore, the homeostatic trafficking, expansion and division of CTV‐labeled congenic donor T cells could be detected using spectral cytometry, in an adoptive T‐cell transfer mouse model. Our optimized CTV method can be applied to both in vitro and in vivo settings to examine the behavior and phenotype of activated T cells.

中文翻译：

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改良的细胞微量紫增殖测定可保留淋巴细胞活力并允许进行光谱流式细胞术分析

在这项研究中，我们描述了用细胞微量紫 (CTV) 标记 T 淋巴细胞的三种不同方法，以便在体外和体内环境中跟踪小鼠和人类细胞的细胞分裂。我们确定了一种改进的 CTV 标记方法，可直接应用于传统或光谱流式细胞术，保持淋巴细胞的活力和功能，同时最大限度地减少染料溢出到其他荧光染料通道中。我们优化的 CTV 标记方法使我们能够识别体外刺激的小鼠和人类淋巴细胞的多达八次细胞分裂和复制指数，以及 T 细胞亚群标记的共表达。此外，在过继性 T 细胞移植小鼠模型中，可以使用光谱细胞术检测 CTV 标记的同源供体 T 细胞的稳态运输、扩增和分裂。我们优化的 CTV 方法可应用于体外和体内环境，以检查活化 T 细胞的行为和表型。