Journal: Journal of Extracellular Vesicles

Article Title: GMP‐Compliant Process for the Manufacturing of an Extracellular Vesicles‐Enriched Secretome Product Derived From Cardiovascular Progenitor Cells Suitable for a Phase I Clinical Trial

doi: 10.1002/jev2.70145

Figure Lengend Snippet: EV enrichment in the final product after purification and concentration by TFF. (a) Particle size distribution. Size distribution profiles were obtained by NTA in spent medium, in CCM after clarification, in the retentate, and in the final product. Data were analysed with the NTA 3.2 software, and the mean and mode of each condition were determined. The data presented are from a single representative experiment. (b) Particle identity and tetraspanin enrichment over the manufacturing process. MFI of CD9, CD63 and CD81 obtained by MACSPlex Exosome kit are presented for spent media, CCM after clarification, retentate and final product. (c) Particle surface marker expression. An abundance of surface proteins on EV in the final product were determined by MACSPlex Exosome kit. (d) Particle concentration. The average concentration (in 10 9 particles/mL) of each sample was determined by NTA and by a correlation between NTA and MACPlex results. (e) Cryo‐EM analysis. Representative images of the final product depicting single‐ and multi‐bilayer bound vesicles. Scale bar = 100 nm. (f) ONi Super‐resolution Nanoparticle imaging. Representative images of a Triple‐Positive (TP) tetraspanin cluster identified within the final product. Individual CD9, CD63 and CD81 images and an overlay are presented. The count (%) for each cluster sub‐type (single, double or triple positive) for the final product is presented in the bar graph.

Article Snippet: A bead‐based multiplex immunoassay, MACSPlex Cytotoxic T/NK Cell Kit (Miltenyi Biotec, #130‐125‐800), was used to measure the levels of multiple cytokines, including Interferon‐gamma (IFN‐γ) and Interleukine‐2 (IL‐2).

Techniques: Purification, Concentration Assay, Clarification Assay, Software, Marker, Expressing, Cryo-EM Sample Prep, Imaging

Journal: Journal of Extracellular Vesicles

Article Title: GMP‐Compliant Process for the Manufacturing of an Extracellular Vesicles‐Enriched Secretome Product Derived From Cardiovascular Progenitor Cells Suitable for a Phase I Clinical Trial

doi: 10.1002/jev2.70145

Figure Lengend Snippet: Long term stability (up to 36 months of storage). (a) Particle size distribution with mean and mode particle size in the final product, by NTA, over time. (b) Tetraspanin marker expression (CD81, CD63 and CD9) in final product by MACSPlex Exosome kit, over time. (c) HUVEC proliferation assay results for final product, over time.

Article Snippet: A bead‐based multiplex immunoassay, MACSPlex Cytotoxic T/NK Cell Kit (Miltenyi Biotec, #130‐125‐800), was used to measure the levels of multiple cytokines, including Interferon‐gamma (IFN‐γ) and Interleukine‐2 (IL‐2).

Techniques: Marker, Expressing, Proliferation Assay

Journal: Journal of Extracellular Vesicles

Article Title: GMP‐Compliant Process for the Manufacturing of an Extracellular Vesicles‐Enriched Secretome Product Derived From Cardiovascular Progenitor Cells Suitable for a Phase I Clinical Trial

doi: 10.1002/jev2.70145

Figure Lengend Snippet: Summary of the components and biological effects of the EV‐enriched secretome final product. (a) Some of the protein and molecular components identified in the final product. Image created with BioRender.com . (b) Summary of cell surface markers identified by the MACSPlex Exosome kit in the final product, which their known roles in biological processes. (c) Functional effects of the final product. Image created with BioRender.com .

Article Snippet: A bead‐based multiplex immunoassay, MACSPlex Cytotoxic T/NK Cell Kit (Miltenyi Biotec, #130‐125‐800), was used to measure the levels of multiple cytokines, including Interferon‐gamma (IFN‐γ) and Interleukine‐2 (IL‐2).

Techniques: Functional Assay