Percoll/Ficoll gradients cause a lot of trouble. Better to use a Miltenyi kit (if you have the magnets) directly on the dissociated cells. Worked fine for us.

Hi Diana,

I am in line with what Kay-Dietrich said above. I have been using Miltenyi microbeads with high purity and efficacy. You can easily isolate many more splenic DCs and NK cells without enrichment. Hope it helps :)

Good luck,

Sudhanshu

I've used this protocol for mouse DC:

Spleen cells were recovered from mice by cutting the

spleen into small fragments and incubating with collagenase D

type II (Roche Biomedical Laboratories, Burlington, NC) solution

(2 mg/ml) for 30 min at 37°C. Digestion was stopped before

collection of cell suspensions. The resulting material was then

passed through a 70-lm cell strainer, and the recovered cells were

subjected to dendritic cell enrichment using magnetic separation

with mouse Pan DC microbeads and a MACS separator (Miltenyi

Biotec).

Hope it can work for you!

Exactly 

Thank you I will try directly the Milteny KIt without enrinchement .