Cryopreservation of Helianthus tuberosus cell suspension cultures: the effect of preculture treatments on cytoskeletal proteins and transglutaminase activity.
Helianthus tuberosus cell suspension cultures were subjected to cryopreservation 24h preculture treatments with 0.5M sucrose or mannitol. Extracts were assayed for transglutaminase activity and the level of alpha-tubulin tyrosination. There was a significant reduction (compared with the non-precultured controls) in transglutaminase activity and alpha-tubulin tyrosination state after mannitol preculture treatment, whereas sucrose preculture treatment produced no significant effect. The results suggest that reduced levels of transglutaminase activity and alpha-tubulin tyrosination are associated with a lack of post-thaw recovery observed following mannitol preculture treatment of cell culture suspensions. These activities may represent useful molecular markers of the success of preculture treatments in cryopreservation protocols.
Harris
W
W
School of Science, The Nottingham Trent University, Clifton Lane, Nottingham NG11 8NS, UK.
Lynch
P T
PT
Hargreave
A J
AJ
Bonner
P L R
PL
D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000008:administration & dosage; D046508:Culture Techniques; D003598:Cytoskeletal Proteins / Q000502:physiology*; D006368:Helianthus / Q000166:cytology* / Q000201:enzymology*; D008353:Mannitol / Q000008:administration & dosage; D008870:Microtubules / Q000502:physiology; D013395:Sucrose / Q000008:administration & dosage; D011503:Transglutaminases / Q000502:physiology*
D003451:Cryoprotective Agents; D003598:Cytoskeletal Proteins; D008353:Mannitol; D013395:Sucrose; D011503:Transglutaminases