Previously described iNGN cells were grown in mTeSR1 medium (Stemcell Technologies) on Matrigel (Corning)-coated plates. Doxycycline (Sigma-Aldrich) was diluted in PBS and added to mTeSR1 medium at a final concentration of 0.5 μg ml−1 to initiate differentiation. On day 4 after Doxycycline addition, the medium was changed to Gibco DMEM with GlutaMAX (Thermo Fisher Scientific), supplemented with B-27 Serum-Free Supplement (Thermo Fisher Scientific) and Gibco penicillin–streptomycin (Thermo Fisher Scientific). On day 6, EVs were isolated by differential ultracentrifugation, as previously described16 (link), and resuspended in PBS.
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Isolation of Extracellular Vesicles from iNGN Cells
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Other organizations : Brigham and Women's Hospital, Harvard University, Massachusetts Institute of Technology, University of Pennsylvania
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Variable analysis
independent variables
- Doxycycline (Sigma-Aldrich) concentration (0.5 μg ml^-1)
- Extracellular Vesicles (EVs) isolated by differential ultracentrifugation
- Previously described iNGN cells
- Matrigel (Corning)-coated plates
- MTeSR1 medium (Stemcell Technologies)
- Gibco DMEM with GlutaMAX (Thermo Fisher Scientific), supplemented with B-27 Serum-Free Supplement (Thermo Fisher Scientific) and Gibco penicillin–streptomycin (Thermo Fisher Scientific)
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