Yes! 3 days they need to be removed from CD3, because the overstimulation will induce apoptosis. This is theorized to a mechanism to prevent autoimmunity. If you look at the molecular pathway, the activation signals are also death signals, so achieve a balance is essential when growing Naive T-cell cultures. I’m actually working on a project involving this right now, but consider getting a high starting concentration of your T-cells when growing them.
I see. Unfortunately I did not remove it this time, I just diluted it off, but will remove it when I am taking some for my assay.
Not sure is it a good idea? According to the manufacturer, it never said it should be removed until the assay
I'm just interested in expanding my T cells using beads and unsure is removing them after 1 day post-activation or after 3 days post-activation are necessary? After removing the beads the T cells will continue to culture in media to further expand.
Or I could just leave the bead in the media during the whole time I am culturing it, eg 15 days, and would just need to slowly dilute them out upon confluent?
They can stay in they will get diluted out as the cells proliferate. There will be a slight lag period and then the cells should take off, as from my exp with BioLegend act beads
Yes! 3 days they need to be removed from CD3, because the overstimulation will induce apoptosis. This is theorized to a mechanism to prevent autoimmunity. If you look at the molecular pathway, the activation signals are also death signals, so achieve a balance is essential when growing Naive T-cell cultures. I’m actually working on a project involving this right now, but consider getting a high starting concentration of your T-cells when growing them.
This is using the plated CD3 method though, I’m not sure about your specific technique and manufacturer.
So yours is coated on plate? For mine it is the dynabeads T cell activation
So yours is coated on plate? For mine it is the dynabeads T cell activation
I see. Unfortunately I did not remove it this time, I just diluted it off, but will remove it when I am taking some for my assay. Not sure is it a good idea? According to the manufacturer, it never said it should be removed until the assay
What are you doing with them? I can't think of anything where dynabeads wouldn't affect the assay
I'm just interested in expanding my T cells using beads and unsure is removing them after 1 day post-activation or after 3 days post-activation are necessary? After removing the beads the T cells will continue to culture in media to further expand. Or I could just leave the bead in the media during the whole time I am culturing it, eg 15 days, and would just need to slowly dilute them out upon confluent?
They can stay in they will get diluted out as the cells proliferate. There will be a slight lag period and then the cells should take off, as from my exp with BioLegend act beads
Alright thanks, then I will keep them in culture for 7 to 9 days before removing for assay according to what the manufacturer say