reezing Whole Bone Marrow Organoids

[Katrine Juhlin Kiørboe]

Hi everyone,  

Has anyone here experimented with freezing whole bone marrow organoids? I’m particularly concerned about the viability of granulocytes post-thaw, as they might not survive the freezing process. If anyone has experience with freezing these organoids, I’d love to hear about the methods you've used and any insights you may have.

Additionally, Abs, I know you’ve already attempted freezing the 'organoids' at an early stage. At which stage in the protocol did you carried this out and how did it worked out for you?

Also, thank you so much for creating this forum—it's wonderful to have a space where we can easily exchange experiences and ideas!  

Best Katrine :) 

[Abdullah Khan]

Hi Katrine, 

Do you mean freezing whole organoids or freezing dissociated organoids? 

For the former - we have tried this with no luck sadly, you just get a lot of cell death post thaw. We've frozen whole organoids at day 14, 18 and 30 I think from memory and nothing really wrks s far. 

We routinely dissociate and freeze the organoids for different assays (e.g. to get repeats in for a 10x run). I have done a head to head comparison and you're right the main population affected by the freezing is mature granulocyte, but otherwise it's pretty comparable to 'fresh' sample. 

If you could clarify what you're after we can advise! Generally if you're dissociating, using something like CryoSTOR and a high cell density will give you a good vial's worth of cells

Thanks! 

Abs