Hi,
I just started working with breast cancer organoids, and currently using 1ml of freezing medium, which contains 10% of DMSO and 90% of FBS (works perfectly well) to make freeze down.
Looking for some more recommendation what medium or reagent to use for freezing organoids
Thanks in advance!
Hello Pravin Phadatare
The ideal freezing medium would be the one that is able to mitigate temperature-induced cell stress responses by scavenging free radicals, pH buffering, providing oncotic/osmotic support, energy substrates, and ionic concentrations that balance the intracellular state at ultralow temperature environments (−80°C to −196°C).
You can also use freezing medium containing (50% FBS, 40% growth media, 10% DMSO).
Gradual freezing to ≤ -80°C using an insulated cryofreezing container is recommended. Organoids should be thawed rapidly in a 37°C water bath and immediately diluted in 10 ml HBSS + 2% FBS per 1 ml freezing media. Centrifuge thawed organoids at 250 rcf for 5 minutes and resuspend in organoid culture media for plating.
Good Luck.
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