Angiogenic Potential of MSC Secretome

Wells of a 96-well plate were coated with 50 μL of Matrigel (Corning) and incubated at 37°C for 30 min to allow Matrigel polymerization. Later, 20,000 HUVECs, re-suspended in the SF medium, were seeded on the top of the Matrigel and incubated at 37°C and 5% CO₂ for 6 h in the presence of different secretome fractions (CM, DM, mEVs, and sEVs) derived from 60,000 MSCs. Images were acquired using a Leica DMi1 inverted phase microscope, and analysis was performed on at least three different areas using the ImageJ Angiogenesis Analyzer plugin (Carpentier et al., 2020 (link)). Data were normalized to a positive control.