PBMCs were collected from whole blood of healthy donors. PBMCs were obtained from buffy coats by Ficoll-Paque (GE Healthcare Biosciences, Piscataway, NJ; catalog 17–1440-02) density gradient centrifugation at 400 g at 20 °C for 40 min to separate blood constituent parts. After purified cells were washed with PBS, CD14+ monocytes were isolated using the Pan Monocyte Isolation kit (Miltenyl Biotec, San Diego, CA; catalog 130–096-537) followed by separation using magnetic LS columns (Miltenyl Biotec; catalog 130–042-401), according to the manufacturer’s protocol.
Macrophages were differentiated from these monocytes according to the previous reports [31 (link), 32 (link)]. The monocytes were cultured in RPMI1640 with either GM-CSF at 20 ng/ml (named M0-GM) or M-CSF at 20 ng/ml (named M0-M) for 5 days. M0-GM macrophages were then stimulated by LPS (20 ng / ml) and IFN-γ (20 ng / ml) for 4 days to polarize into CD80+ / CD86+ (M1) macrophages. On the other hand, M0-M macrophages were stimulated by IL-4 (20 ng / ml) and IL-13 (20 ng / ml) for 4 days to polarize into CD163+ or CD206+ (M2) macrophages (Fig. S3 A-C). M0-GM or M0-M macrophages were stimulated by EVs (20 μg / ml) for 4 days to assess the effects of EVs.
Macrophages were differentiated from these monocytes according to the previous reports [31 (link), 32 (link)]. The monocytes were cultured in RPMI1640 with either GM-CSF at 20 ng/ml (named M0-GM) or M-CSF at 20 ng/ml (named M0-M) for 5 days. M0-GM macrophages were then stimulated by LPS (20 ng / ml) and IFN-γ (20 ng / ml) for 4 days to polarize into CD80+ / CD86+ (M1) macrophages. On the other hand, M0-M macrophages were stimulated by IL-4 (20 ng / ml) and IL-13 (20 ng / ml) for 4 days to polarize into CD163+ or CD206+ (M2) macrophages (Fig. S
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