Witkowski Jacek M
Medical University of Gdańsk, Gdańsk, Poland.
Curr Protoc Cytom. 2008 Apr;Chapter 9:Unit9.25. doi: 10.1002/0471142956.cy0925s44.
This unit proposes a method to extend the already well known dividing-cell-tracking (DCT) cytometric technique based on supravital staining of the lymphocytes with CFSE and allowing them to divide afterwards, beyond simple observation and counting of dividing cells and their generations. Dynamic proliferation parameters that make it possible to determine for in vitro dividing human lymphocytes from various sources, are the actual duration of the pre-division transition period (G0-->G1), time of a single division, and number of divisions an average dividing cell performs over the time of an experiment, as well as the number of effective precursors giving rise to viable daughter lymphocytes. As the method does not require purification of the lymphocyte population of interest, yet allows the calculations for any cytometrically discernible subpopulation, it presents a powerful tool for detailed analysis of the efficiency of proliferative response of the immune cells.
本单元提出了一种方法，该方法基于用CFSE对淋巴细胞进行超活染色并使其随后进行分裂，对已广为人知的分裂细胞追踪（DCT）细胞计数技术进行扩展，而不仅仅是简单地观察和计数分裂细胞及其代数。动态增殖参数使得能够确定来自各种来源的体外分裂的人淋巴细胞的分裂前过渡期（G0→G1）的实际持续时间、单次分裂的时间、平均分裂细胞在实验期间进行的分裂次数，以及产生有活力的子代淋巴细胞的有效前体的数量。由于该方法不需要纯化感兴趣的淋巴细胞群体，但允许对任何通过细胞计数可辨别的亚群进行计算，因此它是详细分析免疫细胞增殖反应效率的有力工具。
