an in vitro model for hepatocyte-like cell differentiation from wharton’s jelly derived-mesenchymal stem cells by cell-base aggregates

cellsderived from wharton’s jelly have been reported to display a wide multilineagedifferentiation potential, showing some similarities to both embryonic (esc)and mesenchymal stem cells (mscs). the present study investigated thedifferentiation potential of human umbilical cord mesenchymal stem cells(ucmscs) into hepatic lineage through embryonic body- like aggregates formationin the presence of igf-i.humanmscs were isolated from the umbilical cord and characterized by flow cytometry;then, their ability to differentiate into osteogenic and adipogenic lineageswas verified. ucmscs were plated in 20 ?l micro drops. after 3 days, 3dspheroids were cultured on the gelatinized culture plates. a two-stepsdifferentiation protocol was used and the cell aggregates were exposed to themedia supplemented with igf, hgf, oncostatin m, and dexamethasone for 21 days.immunoperoxidase and immuno-fluorescence were performed for cyrokeratin 18, 19and albumin. functional assays were done by periodic acid schiff (pas) andindocyanine green.  thecells derived from 3d spheroids showed a polygonal shape after being exposed tohepatogenic media. immunostaining demonstrated the expression of cytokeratin-18,19 and albumin by the differentiated cells. besides, pas staining revealedglycogen storage in differentiated cells. also a greater number of large sizedifferentiated cells were found at the periphery of the expanded cellaggregates.weestablished a protocol for ucmscs differentiation into hepatocytes and thesecells were morphologically and functionally similar to hepatocytes. thus,hepatocyte differentiation may be facilitated by ucmscs aggregate formationbefore administration of the differentiation protocols.