During Cryopreservation, Why DMSO is not suitable for all cell types? And, therefore why glycerol can be used as an alternative?
https://www.researchgate.net/post/Can_we_use_glycerol_to_freeze_cells_instead_of_DMSO
some useful answers
What cell type are you using that cannot handle DMSO? Ethylene glycol or propylene glycol are other alternatives to DMSO or glycerol. Trial and error is the only way to determine the absolute optimal option.
Actually, I am presently planning to work with H9C2 cell line. So, in this case which one I have to used during cryopreservation step? @ Ron Hrstka
I would try the following from Sigma-Aldrich website:
https://www.sigmaaldrich.com/catalog/product/sigma/c9249?lang=en®ion=US.
This is also greener alternative but one has to try for their individual applications.
Hope this helps.
I think, trial and error is the only way to determine the absolute optimal option.
Different cryoprtectants have different molecular sizes and levels of toxicity. Smaller molecule size cryoprotectants like DMSO, glycerol, ethylene glycol etc are called intracellular because they cross the cell membrane, replace internal water to protect from ice crystal damage. Larger molecule size cryoprotectsnts like sucrose can't cross cell membrane, therefore called extracellular. They dehydrate cells by moving water out of the cell.
Different cryoprotectants have differnt safety limits. For example glycerol is less toxic to cells even at higher conc, however, DMSO has narrow safety limit and a little higher conc may become toxic. To overcome this, combination of cryoprotectants is used. For example if DMSO is used along with ethylene glycol, a lower conc of DMSO is needed at which it is not toxic to cells.
Exposure time to cryoprotectant is another big factor that affects the toxicity of cryoprotectants. For example during vitrification, exposure time is less and even higher concentrations are not toxic. Whereas, in slow/long time cryopreservation, much smaller concentrations are used.
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