title: Synchronization of Medicago sativa cell suspension culture.
abstract: Deepening our knowledge on the regulation of the plant cell division cycle depends on techniques that allow for the enrichment of cell populations in defined cell cycle phases. Synchronization of cell division can be achieved using different plant tissues; however, well-established cell suspension cultures provide the largest amount of biological sample for further analysis. Here we describe the methodology of the establishment, propagation, and analysis of a Medicago sativa suspension culture that can be used for efficient synchronization of the cell division and also the application and removal of hydroxyurea blocking agent. A novel method is used for the estimation of cell portion that enters S phase during the assay. The protocol can be used in the case of other species as well.
authors.affiliation: Cellular Imaging Laboratory, Biological Research Centre, Hungarian Academy of Sciences, H-6726, Szeged, Temesvárt krt. 62, Hungary. ferhan@brc.hu
authors.forename: Ferhan
authors.initials: F
authors.lastname: Ayaydin
authors.forename: Edit
authors.initials: E
authors.lastname: Kotogány
authors.forename: Edit
authors.initials: E
authors.lastname: Abrahám
authors.forename: Gábor V
authors.initials: GV
authors.lastname: Horváth
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002453:Cell Cycle / Q000187:drug effects* / Q000235:genetics; D002478:Cells, Cultured; D005434:Flow Cytometry; D006918:Hydroxyurea / Q000494:pharmacology; D000455:Medicago sativa / Q000166:cytology* / Q000187:drug effects*; D008856:Microscopy, Fluorescence; D008940:Mitotic Index; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology*; D013194:Staining and Labeling
chemical_list: D019384:Nucleic Acid Synthesis Inhibitors; D006918:Hydroxyurea