To study the secretome of MSCs and OAd-MSC, 4 × 104 cells per well were seeded in 24-well plates with complete DMEM. After 24 hours, supernatants were collected and proinflammatory cytokines were analyzed using Proteome Profiler Mouse Array Panel A kit (R&D Systems). Cytokine expression was measured semiquantitatively by pixel density of duplicated spots using ImageJ software. Quantification of CXCL10 was confirmed using Mouse CRG-2/IP-10 Ray Bio ELISA Kit (RayBiotech). Differentially expressed proteins were further studied with the STRING (Search Tool for Recurring Instances of Neighbouring Genes) database to analyze the biological process in which they are involved. STRING is a tool that retrieves and displays all known and predicted associations between proteins, including both physical interactions as well as functional associations (19, 20 (link)).
To study the systemic proinflammatory status of mice treated with OAd-MSC, blood samples were acquired 48 hours after treatment administration and serum was obtained following standard protocol. Proinflammatory cytokines were analyzed and measured semiquantitatively as explained above.
To study the systemic proinflammatory status of mice treated with OAd-MSC, blood samples were acquired 48 hours after treatment administration and serum was obtained following standard protocol. Proinflammatory cytokines were analyzed and measured semiquantitatively as explained above.