Journal: Bioengineering & Translational Medicine

Article Title: Mesenchymal stem cell extracellular vesicle vascularization bioactivity and production yield are responsive to cell culture substrate stiffness

doi: 10.1002/btm2.10743

Figure Lengend Snippet: Substrate stiffness influences BM‐MSC EV production and bioactivity. (a) EV production as quantified by EVs per cell from BM‐MSCs seeded on Sylgard 184 PDMS substrates with different base‐to‐crosslinker ratios. EVs used for this data were from 1 day of collection and isolated and counted separately from the conditioned media from the other 2 days. After media collection, cells were trypsinized and counted ( n = 3). (b) After a scratch was induced, HUVECs were treated with BM‐MSC EVs from the different substrates or growth or basal media, and percent gap closure after 20 h was evaluated via microscopy ( n = 3). (c) HUVECs were resuspended in EV treatments or growth or basal endothelial media, seeded in Matrigel‐coated wells, and tube formation after 3–6 h was quantified by the number of loops that had formed ( n = 3). All values expressed as mean ± SD. All data are representative of at least three independent experiments ( n = 3). Statistical significance was determined by ANOVA; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: To measure in vitro angiogenesis, 48‐well plates were coated with 60 μL of growth factor reduced Matrigel (Corning; 356230) and incubated at 37°C for 30 min. P4 HUVECs were then seeded at 35,000 cells/well with either endothelial growth media (PromoCell; C‐22121) with 1% penicillin–streptomycin (positive control), endothelial basal media (negative control), or endothelial basal media (PromoCell; C‐22221) with 0.1% FBS and 1% penicillin–streptomycin with 5E9 EVs/mL.

Techniques: Isolation, Microscopy

Journal: Bioengineering & Translational Medicine

Article Title: Mesenchymal stem cell extracellular vesicle vascularization bioactivity and production yield are responsive to cell culture substrate stiffness

doi: 10.1002/btm2.10743

Figure Lengend Snippet: Softer 184:527 PDMS substrates improve the angiogenic bioactivity of BM‐MSC EVs. (a) EV production quantified as EV per cell from BM‐MSCs seeded on each substrate made with different ratios of Sylgard 184 and Sylgard 527 ( n = 2). EVs used for this data were from 1 day of collection and isolated and counted separately from the conditioned media from the other 2 days. After media collection, cells were trypsinized and counted. (b) After a scratch was induced, HUVECs were treated with BM‐MSC EVs from the different substrates or growth or basal media, and percent gap closure after 20 h was evaluated via microscopy ( n = 3). (c) HUVECs were resuspended in the different EV treatments or growth or basal endothelial basal media, and tube formation after 3–6 h was quantified by the number of loops that had formed ( n = 3). All values expressed as mean ± SD. Statistical significance was determined by ANOVA; * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Article Snippet: To measure in vitro angiogenesis, 48‐well plates were coated with 60 μL of growth factor reduced Matrigel (Corning; 356230) and incubated at 37°C for 30 min. P4 HUVECs were then seeded at 35,000 cells/well with either endothelial growth media (PromoCell; C‐22121) with 1% penicillin–streptomycin (positive control), endothelial basal media (negative control), or endothelial basal media (PromoCell; C‐22221) with 0.1% FBS and 1% penicillin–streptomycin with 5E9 EVs/mL.

Techniques: Isolation, Microscopy

Journal: Clinical and Translational Medicine

Article Title: Targeting capacity, safety and efficacy of engineered extracellular vesicles delivered by transdermal microneedles to treat plasmacytoma in mice

doi: 10.1002/ctm2.70327

Figure Lengend Snippet: CD38‐EVs targeting tumour cells in vitro. PKH26‐labelled extracellular vesicles (EVs) and CD38‐EVs incubated with RPMI8226 cells (A) and U266 cells (B) for 4, 8 and 12 h, with cell fluorescence measured by flow cytometry (FCM). Kinetics of uptake of EVs and CD38‐EVs within 12 h by RPMI8226 cells (C) and U266 cells (D), measured by relative fluorescence units (RFU). (E) RPMI8226 and U266 cells were incubated with EVs and CD38‐EVs for 8 h and visualised using laser scanning confocal microscope (LSCM) (scale bars: 5 µm). PKH26‐labelled EVs and CD38‐EVs incubated with RPMI8226/HUVECs and RPMI8226/BMSCs (F) and U266/HUVECs and U266/BMSCs (G) co‐culture systems for 12 h, with cell fluorescence measured by FCM. Kinetics of uptake of EVs and CD38‐EVs by myeloma cells, human umbilical vein endothelial cells (HUVECs) and bone marrow stromal/stem cells (BMSCs) in RPMI8226/HUVECs and RPMI8226/BMSCs (H) and U266/HUVECs and U266/BMSCs (I) co‐culture systems, measured by RFU. Data are generated from three independent experiments.

Article Snippet: The HUVECs cell line was grown in ECM medium containing an endothelial cell growth medium bullet kit (Sciencell).

Techniques: In Vitro, Incubation, Fluorescence, Flow Cytometry, Microscopy, Co-Culture Assay, Generated