Dec 15, 2023
Patient PBMC differentiation to macrophages
- 1University of Florida
Protocol Citation: Rebecca Wallings 2023. Patient PBMC differentiation to macrophages. protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr3w83vmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: December 14, 2023
Last Modified: December 15, 2023
Protocol Integer ID: 92334
Funders Acknowledgement:
ASAP
Grant ID: ASAP-020527
Abstract
Patient PBMC differentiation to macrophages
After cryorecovery, plate PBMCs at a final concentration of 3 x 106/mL in a poly-D-lysine coated, 96-well plate, resulting in 3x105 cells per well.
Incubate PBMCs at 37°C for 2-hours to allow adherent cells to adhere.
Wash cells washed 2 x sterile PBS to wash off non-adherent cells, and incubate in growth media
containing 20ng/mL of human macrophage-colony stimulating factor (hM-CSF) for 5 days, with a full media change on day 3 to replace nutrients.