Fig. 5

The optimization and in vitro characterization of docetaxel-loaded nanovesicles. a iPSC-MSCs were not pretreated or pretreated with 5 μg/mL Dxl for 24 h, and then broken down by serial extrusion in 50, 100, or 200 μg/mL Dxl to make Dxl-NVs. Dxl-NVs were isolated by ultra-centrifugation and the Dxl loading in NVs was measured by UV spectrometry. b Sizes of empty NVs or Dxl-NVs extruded in 50, 100, or 200 μg/mL Dxl (NV 50, NV 100, NV 200) were determined by Nanosight nanoparticle tracking analyses. c, d Effects of Dxl loading on selective uptake of DiI-labeled NVs by PC3 cells vs. SMCs were examined by flow cytometry and the LORs were calculated as in Fig. 1. e NV-Dxl was incubated in 37 °C PBS containing 10% human serum or 4 °C PBS for a series of periods, and the supernatant was isolated by ultra-centrifugation to measure Dxl release by UV spectrometry. Parent PC3 cells (f), Dxl-resistant PC3 cells (g), or THP-1 human myeloid cells (h) were incubated with empty NVs, NV-Dxl, or free Dxl at a serial of concentrations for 72 h and then analyzed with PrestoBlue Cell Viability Assay (ThermoFisher). N = 3 in all assays, *p < 0.05 vs. all other groups