I am interested in the study of dendritic cells response under infection with mutant strain candidate to vaccine against TB.
But I am begginer in the work with dendritic cells, somebody can help me?
Thanks
Thanks very much, I am reading the manuscript
¿Do you know ther percentage of DC from MN?
Greetings
Hello Maria Teresa,
We have never measured that quantitatively. I've also been retired for 10 years now. We used human donors from a blood bank to obtain DC from monocytes. So there were slight differences from donor to donor. Occasionally CD14 remained too high, so we set ourselves a limit on what we would tolerate. Some of the cells also die during differentiation. If I remember correctly, about 70 - 80 % of the seeded monocytes went into the experiment as DC. In exceptional cases, however, even less
Greetings
Jochem
Thanks very much for your time and your expertise.
For to characterized DC, what markers do you recomended? for fluocytometry
Hello Maria Teresa,
when we started the test we determined CD14, CD1a, CD86, CD80 and HLA-DR. CD14 and CD1a to check how far the monocytes had developed into DC and CD86 and CD80 as markers for the reaction to potential allergens. We did not test CD80 later as it turned out that the AK CD80 from different manufacturers gave completely different results. We changed the AK's once as we had received a new FACS device. Later we only did a follow-up with CD14 and CD1a and for the actual detection of the extent to which the cells reacted to the substances we used CD86. We tested the responsiveness of the cells with LPS.
Greetings
Jochem
Hi Maria Theresa,
I don't know if it's helpful, but a few details about our protocol
Jochem
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