Symposium “Plants in Health and Culture”
L. Suhartono1, F. van Iren2, Y. H. Choi2 and R. Verpoortel
1Department of Pharmacognosy, Institute of Biology, Section of Metabolomics, Leiden University, Leiden, The Netherlands
2Institute of Molecular Plant Sciences, Leiden University, Leiden, The Netherlands
Cryopreservation and metabolite profiling of suspension cultured Nicotiana tabacum cv bright yellow-2 cells
The metabolite profile of Nicotiana tabacum (BY-2) suspension cell culture before and after cryopreservation was studied by metabolomics approach. For the cryopreservation procedure, BY-2 cells were precultured in LS media supplemented with mannitol for 3 days following with incubation in LS media supplemented with 1M sucrose, 0.5M glycerol, 0.5M DMSO and 0.04M L-proline as cryoprotectants in an ice bath for 20 min, slow cooling for 4 hours in a -80 °C freezer followed by rapid immersion in liquid nitrogen, rapid thawing and transfer of the cells without washing to LS media solidified with 7% agarose. After two weeks of recovery, the cells were transferred into liquid medium. 1H NMR and multi variate analysis (PCA) were performed to analyse the metabolites of BY-2 cells.