What is the type of cells you want to culture from PBMC exactly?

Normally, monocytes and thrombocytes will adhere quickly within 30 minutes incubation at 37C, while lymhocytes will float. So if you see floating cells from your PBMC culture, they are definitely lymphocytes. If you don't need them just wash them away.

Monocytes will adhere in standard cell culture plate coated with poly-l-lysine or collagen.

The amount of cells you can put in a well depends on the size. For bigger cells such as fibroblast, I usually put 20-30 thousands/well, for medium size cells such as A549, HeLa, I put up to 50 thousands/well. Smaller cells like lymphocytes or 293 cells, I put up to 80 thousands. It also depend on your objective, because you may want to use exact number of cells for your experiment rather than confluency.

Following

Only DCs and macrophages are expected to adhere on the tissue culture plates. Generally, in mouse PBMCs you may find less than 10% of these populations.

There are tissue culture treated 96 well plates available commercially. We buy corning products however, there may be other sources as well.

If your objective is to obtain large quantities of macrophages then peritoneal-exudate-cells derived macrophages may be better option.

You may also get a lot of macrophage DCs from bone marrow.

Good luck

Sudeep

Hello

I agree with Quick Ida ; Following article may be helpful for you

https://bio-protocol.org/e322

Good Luck