Figure 2
Characterization of eMSCs (a) and eECs (b). (A) The outgrowth of the mix population from the explanted tissue under MSC medium condition. (B) The eMSCs had a spindle-shaped morphology, and (D) positive Vimentin staining indicated the presence of this intermediate filament protein (red: nuclei in blue). (C) MSC-specific markers (CD29, 40, 90) were demonstrated by gene expression analysis. (E) As indicators of osteogenic potential, eMSCs exhibited positive ALP staining, and (F) positive alizarin red staining for extracellular calcium deposits under osteogenic culture conditions, confirming differentiation into osteogenic lineages. (G) Oil Red O staining emphasized the presence of intracellular lipid droplets, confirming adipogenic differentiation capability of the eMSCs. The characterization of eECs (b) revealed (H,K) The outgrowth of the epithelial cells from the explanted tissue under FBS-EC and KSR-EC medium condition respectively (I,N) a polygonal cell shape in both EC media tested (FBS-EC and KSR-EC, respectively). (J,O) Positive Pan Cytokeratin staining of the cytoskeleton was evident in eECs in both media. In FBS-EC media, eEC purity, determined by flow cytometry, was 94.78% (K), whereas in KSR media, purity reached 97.70% (P). (L,Q) PCR validated the gene expression for the eEC-specific marker Muc1 in eECs derived from both EC media. (Uncropped conventional PCR gels were shown in the Supplementary Uncropped Fig. 1A–F for eMSCs and Figure for eECs).
