Vaz-de-Lima Beatriz Brandão, Ionta Marisa, Machado-Santelli Glaucia M
Universidade de São Paulo, Instituto de Ciências Biomédicas, Departamento de Biologia Celular e do Desenvolvimento, Sao Paulo, Brazil.
Micron. 2008 Jul;39(5):631-4. doi: 10.1016/j.micron.2007.04.007. Epub 2007 May 3.
The scrape loading and dye transfer (SL/DT) method is a functional assay widely used to evaluate the level of the intercellular communication via gap junction. Initially, the evaluation of the degree of communication was done by counting the rows of fluorescent cells from the scrape. Its substitution by fluorescence distance or area, determinated by morphometric softwares, was proposed as the best way of communication measurement. Considering that normal cells change their morphology during different phases of growth in monolayer cultures we proposed to evaluate the possible influence of the cell morphology changes on the quantification of intercellular communication when the SL/DT method is used. The present data indicates that GJIC capacity should be rather quantified by counting the fluorescent cell rows than by the measurement of the fluorescence distance or area, when the goal is to evaluate the cell communication along different phases of cell culture. Normal cells growing in monolayer culture decrease their diameter according to their final density of saturation, impairing the quantification of GJIC capacity by morphometric analysis.
刮擦加载与染料转移（SL/DT）方法是一种广泛用于评估通过间隙连接进行细胞间通讯水平的功能分析方法。最初，通讯程度的评估是通过计数刮擦处荧光细胞的行数来完成的。有人提出用形态测量软件确定的荧光距离或面积来替代它，认为这是测量通讯的最佳方法。考虑到正常细胞在单层培养的不同生长阶段会改变其形态，我们提出评估当使用SL/DT方法时细胞形态变化对细胞间通讯定量的可能影响。目前的数据表明，当目标是评估细胞培养不同阶段的细胞通讯时，间隙连接细胞间通讯（GJIC）能力应该通过计数荧光细胞行来定量，而不是通过测量荧光距离或面积。在单层培养中生长的正常细胞会根据其最终饱和密度减小其直径，这会损害通过形态分析对GJIC能力的定量。
