Human monocyte derived dendritic cells, through in vitro GM-CSF + IL-4 differentiation, are loosely- and non-adherent at their immature state. In my hands, maturation of such dendritic cells (LPS, IFN-g, TNF-a or other stimuli) makes them more adherent. There are many ways to harvest the mature dendritic cells from the culture plate: Pipetting, EDTA, cell scraber, only collecting non-adherent fraction etc.
So, my question is: How do you harvest your mature human monocyte derived dendritic cells?
And, why do you believe that's the correct way?
Hello Esben,
due to the behavior of the dendritic cells as described by you, we have examined in our laboratory both the adherent and the loosen cells separately for CD14, CD1a and CD86. there were no differences between the cells. Therefore, we suspected that the states alternate between loosely and adherent cells. In other words, the cells do not remain permanently adherent, but the state changes. Therefore, when harvesting or transferring the cells, we only rinsed them off and dispensed with the adherent cells, which were present in the minority anyway, insofar as they remained adherent.
Greetings
Jochem
There are multiple methods you can use to harvest mature human monocyte-derived dendritic cells (DCs) from culture plates. Here are some commonly employed techniques:
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