There is a protocol by Menges and Murray (2004) Plant J 37, 635-644, but the main description is for Arabidopsis cells, and I cannot make out the exact protocol used for the BY-2 cells. Does anyone (1) have experience of this protocol for BY-2, (2) know the more specified protocol for the BY-2, or (3) know a different protocol for specifically cryopreserving BY-2? Also, is liquid nitrogen storage essential or can one store the cells in a -150°C freezer?
Hi
Check this protocole for TBY-2 cell cryopreservation in Reinhoud et al. 1995. Vitrification and a heat-shock treatment improve cryopreservation of tobacco cell suspensions compared to two-step freezing.
(30 min. at -35°c then use nitrogen liquid).
It is well recommended to preserve TBY-2 cells that are in their early stationary phase because they are more tolerant to freezing.
Regards
Mohammad
Use 10% dmso with 90% FBS. Store 10 lakh cells in one vial minium or max 15 lakh. No liq N2 not required , store at -20 or -80 degree is best. Liq N2 for long term storage. -80 can help you for 6 months or maximum year. If you want to preserve for long term you need liq N2. Alternative is thaw cell after some time again store after 2-3 passage. Good luck
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