Hi all,
I'm growing primary basal cells from lung digests, and was wondering if anyone has recommendations for cell freezing media for primary lung basal cells.
Previously I have been using Cryostor CS10, however I'm finding quite costly and would like to make my own.
My expansion media is Pneumacult EX Plus.
Many thanks,
Sam
You might try this method with cell culture grade DMSO. I would verify this works with your cells before committing entirely. Good luck!
Article Freezing Medium Containing 5% DMSO Enhances the Cell Viabili...
Thank you Susan!
I have since found another paper that uses 10% DMSO and 30% FCS in their lung basal media which I will also try.
Not all papers report their cryopreservation protocol!
we used 5% cell culture grade DMSO in FBS for cell storage. A higher concentration of DMSO could be toxic to the cells.
Thank you,
Manzoor Ahmed, how much FBS do you use? Or are you using 5% DMSO in undiluted FBS?
Hi Sam,
The UNC MLI Tissue Procurement and Cell Culture Core recommends the use of 5% DMSO for Primary Human Bronchial Epithelial Cells (https://www.med.unc.edu/mlicellcore/research-applications/). I recommend to avoid using FBS since your cells are maintained in a serum-free medium, this could affect their phenotype. You could simply try and add 5% DMSO to PneumaCult-Ex Plus medium.
Martine
Thanks Martine Bail
That actually makes a lot of sense. I was feeling uneasy about adding FCS to a media specially formulated to be serum free! Especially as the FCS will most definitely affect phenotype!
Sam
My pleasure! Let us know how your cells recover from freeze-thaw and how it compares to the current freezing medium!
Cheers,
Martine
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