Hi
I'm trying to activate my T-cells with Dynabeads at a 3:1 ratio. One thing I've noticed is the tendency for the beads to not spread homogenous, whether its in the middle of the well or side of it. I've tried throughly mixing but over time I see that happening.
Is there anything wrong with this? Do I need to get rid of it?
Does this affect the T-cell activation? How do you control that in a clinical setting?
It is normal for the beads to clump and not be spread apart. If using flat bottom wells, both cells and beads tend to aggregate at the center and around the edges and there is not much you can do about it. T cells like to form clusters and grow better when using a round bottom well/tube. Beads also would cluster at the center, thereby providing better contact.
Julie, I am observing that after debead my cells I am not getting consistence in my recovery, but in the last experiments my yield drop more than 60%. I would like to know if you can see some reason for this behaving, once the protocol is the same all the time. PS: do you thing that the concentration of the dynabeads suspension could be influence the recovery? and if in excess the viability of my cell could drop after debead them? Cheer, Fernando.
@julie - thats absolutely correct. check out this video demonstrating bead-cell interaction over time. https://www.youtube.com/watch?v=hDDvgh9F_Dc
Feel free to get in touch at [email protected]
kind regards ketil
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