We are finding difficulties in neutrophils activating and clumping once resuspended either from a fresh sample or a frozen/thawed sample. Sample is acquired from human blood using Sepmate tube and gradient purification.
No calcium or magnesium is being introduced to the cells at any point during the purification process and current buffer being used is 0.5% BSA in PBS + 2 mM EDTA.
Sample is being thawed at 37C and washed in buffer at the sample temperature prior to being centrifuged at 300g for 5 minutes.
I'm grateful for any help, thank you!
Hi Charles, My name is Eilysh and I am a Product Manager at STEMCELL Technologies. Thank you for your question! Please refer to our guide on how to thaw frozen primary cells: https://www.stemcell.com/technical-resources/how-to-thaw-frozen-primary-cells.html. Additionally, we do sell frozen neutrophils. They can be successfully frozen and thawed and then used for downstream assay but their lifespan is limited (less than 6 hours). If you want to use frozen neutrophils, it needs to be immediately after thawing and for something short. Ideally, we would recommend sticking to freshly isolated neutrophils if possible. I hope this helps! If you have any more questions, feel free to email us at [email protected]. Kind regards, Eilysh
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