Hello everyone,

So I am trying to isolate dendritic cells from human PBMCs. I start with a buffycoat of blood from which I isolate the mononucleated cells.

I then do a selection of CD14+ cells with CD14 beads.

In the end I have approx 180 Millions of CD14+ monocytes.

I plate them in 6 well plates with 2 millions cells per mL and 3mL per well.

The media I use is RPMI + 10%FBS and B-mercaptoethanol and 25ng/mL of GMCSF and IL4.

At day 3 i add again GMCSF and IL4 (still the same concentration)

at day 6 I harvest my cells. From 180Millions monocytes I only obtain 35 Million Dendritic cells.

I understood that the yield was supposed to be of 80%.

Does anyone has any idea on how can I increase this yield ? I use the Mitlenyi research grade ones.

Thanks a lot !

Best regards,

Sonia