TABLE 2.
Ex vivo skin mononuclear phagocyte culture optimization
| Parameter | IL−4 | GM−CSF | FCM (%) | Fold change in cell viability | % live |
|---|---|---|---|---|---|
| cDC2 | − | − | − | 1.0 | 23 |
| + | − | − | 0.5 | 11 | |
| − | + | − | 0.9 | 21 | |
| + | + | − | 1.1 | 25 | |
| − | − | 20 | 1.4 | 32 | |
| − | − | 40 | 1.1 | 25 | |
| CD14 monocyte–derived macrophages | − | − | − | 1.0 | 13 |
| + | − | − | 3.6 | 47 | |
| − | + | − | 1.6 | 21 | |
| + | + | − | 2.7 | 35 | |
| − | − | 20 | 3.3 | 43 | |
| − | − | 40 | 2.2 | 29 | |
| AF+ macrophages | − | − | − | 1.0 | 12 |
| + | − | − | 1.0 | 12 | |
| − | + | − | 0.3 | 3.6 | |
| + | + | − | 1.1 | 13 | |
| − | − | 20 | 4.9 | 59 | |
| − | − | 40 | 2.6 | 31 |
Human skin mononuclear phagocytes were isolated from human skin with type IV collagenase, were sorted by FACS, and then cultured for 96 h in 96-well, round-bottom plates at 2 × 105 cells/well under various conditions. The percentage of live cells was then calculated using live/dead near-infrared staining and true count beads (n = 3). AF, autofluorescence; FCM, fibroblast conditioned media.