E‐cadherin synergistically induces hepatospecific phenotype and maturation of embryonic stem cells in conjunction with hepatotrophic factors

Abstract

Since effective cell sourcing is a major challenge for the therapeutic management of liver disease and liver failure, embryonic stem (ES) cells are being widely investigated as a promising source of hepatic-like cells with their proliferative and pluripotent capacities. Cellcell interactions are crucial in embryonic development modulating adhesive and signaling functions; specifically, the cell-cell adhesion ligand, cadherin is instrumental in gastrulation and hepatic morphogenesis. Inspired by the role of cadherins in development, we investigated the role of expression of E-cadherin in cultured murine ES cells on the induction of hepatospecific phenotype and maturation. The cadherin-expressing embryonic stem (CE-ES) cells intrinsically formed pronounced cell aggregates and cuboidal morphology whereas cadherin-deficient cadherin-expressing embryonic stem (CD-ES) cells remained more spread out and corded in morphology. Through controlled stimulation with single or combined forms of hepatotrophic growth factors; hepatocyte growth factor (HGF), dexamethasone (DEX) and oncostatin M (OSM), we investigated the progressive maturation of CE-ES cells, in relation to the control, CD-ES cells. Upon growth factor treatment, the CE-ES cells adopted a more compacted morphology, which exhibited a significant hepatocyte-like cuboidal appearance in the presence of DEX-OSM-HGF. In contrast, the CD-ES cells exhibited a mixed morphology and appeared to be more elongated in the presence of DEX-OSM-HGF. Reverse-transcriptase polymerase chain reaction was used to delineate the most differentiating condition in terms of early (alpha-fetoprotein (AFP)), mid (albumin), and late-hepatic (glucose-6-phosphatase) markers in relation to growth factor presentation for both CE-ES and CD-ES cells. We report that following the most differentiating condition of DEX-OSM-HGF stimulation, CE-ES cells expressed increased levels of albumin and glucose-6-phosphatase, whereas the CD-ES cells showed low levels of AFP and marginal levels of albumin and glucose-6-phosphatase. These trends suggest that the membrane expression of E-cadherin in ES cells can elicit a marked response to growth factor stimulation and lead to the induction of later stages of hepatocytic maturation. Thus, cadherin-engineered ES cells could be used to harness the cross-talk between the hepatotrophic and cadherin-based signaling pathways for controlled acceleration of ES hepatodifferentiation.

Control of hepatic differentiation via cellular aggregation in an alginate microenvironment

“…However we do see E-cadherin production later on in the differentiation process, Figure 2A. The E-cadherin protein itself has been identified within liver tissue samples, (Figarella-Branger et al, 1995) and has been reported to play a role in hepatic differentiation, (Brieva and Moghe, 2004;Dasgupta et al, 2005). Furthermore, the presence of tight junctions facilitated by the presence of E-cadherin, are necessary for normal liver function.…”

Enhanced Differentiation of Adult Bone Marrow-Derived Stem Cells to Liver Lineage in Aggregate Culture

“…49 Similarly, mouse ESCs grown as EBs overexpressing E-cadherin were shown to have higher transcript levels of G6pc in response to growth factor cocktails than EBs in which E-cadherin expression was downregulated. 50 Other than cell-cell interactions, polarization of cells in 3D configurations has been shown to help in enhancing and stabilizing the differentiated functions of hepatoyctes. 47,51 Whether similar positive effects of 3D culture on hepatocytes are in play in enhancing differentiation of stem cell certainly requires further investigation.…”

Artificial extracellular matrix for embryonic stem cell cultures: a new frontier of nanobiomaterials

“…The basic idea behind the EB protocol is to mimic the process of embryonic development [13]. Most cell lineages, including the neuronal, mesoderm and hepatocyte lineages have been shown to be generated within the EB [32][33][34]. However, the structure of the EB is far more heterogeneous than the actual embryo.…”