Cell cycle synchronization of tobacco BY-2 cells
| Autor: | Seiichiro Hasezawa, Tomomi Hayashi, Fumi Kumagai-Sano, Toshio Sano |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Tobacco BY-2 cells
Aphidicolin education.field_of_study DNA Plant Cell Population Cell Cycle DNA replication Cell Culture Techniques food and beverages Cell cycle Biology General Biochemistry Genetics and Molecular Biology Cell biology Cell Line chemistry.chemical_compound medicine.anatomical_structure chemistry Tobacco medicine Cell synchronization education Mitosis |
| Zdroj: | Nature protocols. 1(6) |
| ISSN: | 1750-2799 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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