DETERMINATION OF CELL NUMBER IN MONOLAYER-CULTURES
DETERMINATION OF CELL NUMBER IN MONOLAYER-CULTURES
复制标题
DOI：
10.1016/0003-2697(86)90314-3
复制
发表时间：
1986-11-15
影响因子：
2.9
通讯作者：
DENTON, M
中科院分区：
文献类型：
--
作者：
GILLIES, RJ;DIDIER, N;DENTON, M
确定各种条件对单层细胞的细胞生长抑制或细胞毒性作用需要快速、可重现且能够监测这些作用随时间变化的技术。目前用于监测细胞抑制或细胞毒性的方法是静态或间接的;也就是说，它们被设计用于测试在单个时间点或对相关细胞过程（如膜完整性）的各种处理的影响。由于这些限制，在现存的技术，我们进行了这项研究，以改善方法快速测定细胞数量在单层培养。我们已经达到了在固定方案下用结晶紫染色细胞核的条件，该方案允许在24孔微孔中生长的培养物中快速且可重复地定量细胞数量。通过将吸收的染料溶解到Triton-X-100溶液中并测定光密度（O.D.）使用分光光度法。本来文证明，O.D.与细胞数线性相关，灵敏度为CA。500个细胞，并且该技术适用于研究影响细胞增殖的试剂。
Determining the cytostatic or cytotoxic effects of various conditions on monolayer cells requires techniques that are rapid, reproducible, and able to monitor these effects as a function of time. Methods currently used to monitor cytostasis or cytotoxicity are either static or indirect; that is, they are designed to test effects of various treatments either at single time points or on associated cellular processes, such as membrane integrity. Because of these limitations in extant techniques, we undertook this study to improve methods for the rapid determination of cell number in monolayer cultures. We have arrived at conditions of staining cell nuclei with crystal violet under fixed regimens which allow rapid and reproducible quantification of cell number in cultures grown in 24-well miniwells. Quantification is possible by solubilizing the absorbed dye into a solution of Triton-X-100 and determining optical density (O.D.) using spectrophotometry. The present communication documents that O.D. is linearly related to cell number with a sensitivity of ca. 500 cells and that the technique is applicable to study agents which affect cell proliferation.