3.3 MicroRNA profiles of EVs during expansion and
differentiation
Among the variety of
molecules
that are contained in EVs, miRNAs have attracted the
most attention
because of their
regulatory roles in gene
expression [45]. For this reason, we determined the content of
miRNAs within our EVs. We then moved onto next-generation sequencing to
determine the miRNA profiles contained within our EVs. A total of 306
miRNAs were identified in all samples (Table S1) . Clustering
analysis of the miRNA expression in the six groups of EVs revealed
marked differences in miRNA expression, and the software system was able
to determine biological pathways that were significantly impacted by the
identified miRNAs (Figure 4A and Table S2) . Among the
differentially expressed miRNAs in D0 versus D21 (Table S3),miR-186, miR-210, miR-181c-5p, and miR-146a-5p were increased two-fold
and miR-133 and miR-485 were decreased two-fold in the last stage.
Meanwhile, miR-10, miR-27a, miR-192 increased which have been reported
as late markers of osteogenesis, and miR-31 decreased. In addition,
miR-10 and miR-138 were expressed six-fold higher in BMSCs versus PSCs(Figure 4B, C) . The top 34 KEGG pathways were also reported
which indicated the potential upregulated target genes of the 301 miRNAs
that were identified in both BMSC and PSC EVs (Figure S1) . Of
the potential KEGG pathways analyzed Wnt signaling was of particular
interest due to its role in osteogenic differentiation of MSCs [46].