We recently received some Plasmodium falciparum sporozoites cryovials which are frozen with HES (hydroxy ethyl starch),Trehalose and DOPC (Dioleoylphosphatidyl-Choline) based freezing solution. As we have no experience with these freezing solution, can I please ask if anyone can advice us what principle should we follow when we thaw these cells (also would be fantastic if anyone can provide us a thawing protocol for similar conditions)?
I do not know your application but I have used the trehalose based freezing solutions with human epithelial cells. We thawed the cells the same way with any cryo-media, Fast. Threhalose is not considered toxic. Similar to raffinose or other branched carbohydrates.
General procedure:
1. Vials were swirled by hand in a 38 C water bath.
2. As soon as the lat remnant of ice disappears, we very gently diluted the crryo-media 10 fold
3. cells allowed the cells to rest for 30mins-1 hour in aerobic conditions.
The thaw is the highest stress point for cells in the cryopreservation procedure. Respiration restarts and membranes have to be repaired. Between the damaged membranes and the superoxide generation, not all cells will make it out of cryo.
Hope this helps
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