miRNA profiles of EVs released from human umbilical cord mesenchymal stem cells (hUCMSC-EVs)

To identify specific miRNAs carried by hUCMSC-EVs, and explore their crucial roles in hUCMSC-EV-based improvement of inflammatory diseases. miRNAs expression in hUCMSC-EVs derived from two different donors. EVs were isolated from cell-free conditioned medium after 48 hrs of culture using differential ultracentrifugation. Total RNA was extracted with TRIZOL reagent, quantified spectrophotometrically (NanoDrop ND-2000, Thermo Scientific), and assessed for integrity using an Agilent Bioanalyzer 2100 (Agilent Technologies). Following manufacturer protocols, RNA samples were dephosphorylated, denatured, and labeled with Cyanine-3-CTP. After purification, labeled RNA was hybridized to a miRNA microarray. Slides were washed and scanned using the Agilent Scanner G2505C (Agilent Technologies).