Hi!
Right now I am working with embyioid bodies generated from iPSCs (PBMC) to obtain CD34+ cells.
After 2 weekes of culture flow cytometry has to be performed.
It is highly problematic because I can't easily dissociate EBs and maintain high viability of cells.
According to the protocol I used TryPLE Select and use 21G syringe. (source: Iriguchi 2019)
This step is fine but then when I resuspend cells in Binding Buffer for Flow Cytometry, cells become to be highly adherent and they even stick to the tips or needle, they are making clots.
Is there any possibility to dissociate EBs into real single-cell suspension for further analysis?
Thank you for your help!
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