By now, culturing monocytes to create myeloid dendritic cells should be old hat. The technique was first published by Sallusto and Lanzavecchia in 1994. In spite of the widespread use of this culture method, there are still questions about the best practices, time in culture, maturation, and other variables.
We recently received a few questions from customers about this very topic. You may have been wondering the same.
“Do dendritic cells start dying immediately, or will viability be >90% for the first few days, followed by a dropoff?”
“Would culturing dendritic cells in plates that assist adherence (like poly-lysine or gelatin) increase viability/length of culture?”
“Does activation of dendritic cells change viability dynamics?”
“When culturing dendritic cells with serum media, would you recommend heat inactivated or non-heat inactivated serum?”
Answering Your Dendritic Cell Questions
Dendritic Cells Don’t Proliferate
Expect very little to no proliferation in a dendritic cell culture.
While we have detected CD34+ cells in monocyte preparations (which do have proliferation potential) the monocytes themselves are not likely to proliferate.
We don’t recommend culturing human dendritic cells from Cellero unless you’re simply trying to simulate them.
Survival Time in Culture is Short, But It Depends
To be fair, we haven’t formally studied the limit of keeping dendritic cells alive in culture.
We have studied the stimulation of immature dendritic cells with TNF, LPS, poly I:C, and other factors and have found that cell death typically occurs within a few days.
Dendritic Cells are Sticky!
It’s a troublesome feature of dendritic cells (and monocytes and macrophages) that they are tightly adherent to tissue culture plastic. You’ve likely encountered this issue before! The cells stick even tighter when activated by TLR agonists.
Try an ultra low adhesion culture vessel to make your life much easier. But beware, these vessels are expensive. Instead, plan ahead so you can plate the cells where you intend for them to stay.
Experimenting with Dendritic Cells?
Follow our detailed dendritic cell culture protocol and access collection and culture tips from our scientific team.
We also have several lots of monocyte-derived dendritic cells ready to ship when you need them.