I have a line of healthy human astrocytes, and plan to passage it.
I was wondering if any suggestions on how to cryopreserve it?
I use mFreSR from stem cell to freeze the iPSCs and not sure if I can use that for astrocytes too?
I saw a post with comments suggesting 10%DMSO and 90%FBS, but was wondering if the FBS will have any negative effects on human astrocytes and if I need to use a different serum instead, or if washing the cells after thawing will be enough to remove the FBS and limit it's effects.
Thank you,
Mahsa
Hi Mahsa Khayat-Khoei . 10% DMSO, 90% FBS is the standard freezing solution used for all mammalian cells. It is not perfect, but it works. Even mFreSR has DMSO in it.
The serum is in the freezing solution to further protect cells against damaging ice crystal formation. FBS is used because it is the most common serum in labs doing TC. Any serum will do just as well. Anyway, FBS can be washed away upon thawing the cells.
If you plan on banking a lot of cells it might be worth trying to optimize your freezing protocol. Try freezing several vials with different protocols including:
- your existing protocol
- 10% DMSO + 90% serum (ie FBS)
- commercial cryopreservation media
- 10% DMSO + 40% culture media + 50% FBS
- 10% glycerol + 40% culture media + 50% FBS
Then thaw vials, test viability with Trypan blue and chose the method which gives you the best viability.
10%DMSO +90% FBS is pretty good for any types of mammalian cells.
Hi Mahsa Khayat-Khoei
Did you find in the end a good protocol? I'm in the same situation, wondering if FBS may induce negative effects on hiPSC-derived astrocytes.
Thank you!
Martina
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