Protocol abstract
Nature Protocols 1, - 2621 - 2627 (2007)
Published online: 11 January 2007 | doi:10.1038/nprot.2006.381
Subject Categories: Cell and tissue culture | Plant biology
Cell cycle synchronization of tobacco BY-2 cells
Fumi Kumagai-Sano1, Tomomi Hayashi2, Toshio Sano2 & Seiichiro Hasezawa2
Abstract
Synchronization is a powerful technique for understanding cell cycle events. Here, we describe the procedure for synchronizing tobacco bright yellow 2 (BY-2) cell line, with which an exceptionally high level of synchrony can be achieved. It basically relies on an "arrest-and-release" strategy using aphidicolin, an inhibitor of DNA replication, and propyzamide, a plant-microtubule disruptant. In a single-step process using aphidicolin alone, a cell population with about 70% of the cells at mitosis can be achieved, whereas by a two-step method using the two inhibitors sequentially, the level of synchrony can reach over 90%. The method of choice depends not only on the peak mitotic cell proportion but also on the cell cycle stage that is targeted for analysis. Both procedures take about 1.5 days, and cell cycle progression can be observed from the S phase to the next G1 phase at about 12 h after a 24 h-period treatment with aphidicolin.
- Department of Science Education, Faculty of Education, Gunma University, Aramaki-cho 4-2, Maebashi, Gunma 371-8510, Japan.
- Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwanoha 5-1-5, Kashiwa, Chiba 277-8562, Japan.
Correspondence to: Fumi Kumagai-Sano1 e-mail: fsano@edu.gunma-u.ac.jp
