Hyaluronan is Crucial for Stem Cell Differentiation into Smooth Muscle Lineage

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Deciphering the extracellular signals that regulate SMC differentiation from stem cells is vital to further our understanding of the pathogenesis of vascular disease and for development of cell-based therapies and tissue engineering. Hyaluronan (HA) has emerged as an important component of the stem cell niche, however its role during stem cell differentiation is a complicated and inadequately defined process. This study aimed to investigate the role of HA in embryonic stem cell (ESC) differentiation toward a SMC lineage. ESCs were seeded on collagen-IV in differentiation medium to generate ESC-derived SMCs (esSMCs). Differentiation coincided with increased HA synthase (HAS) 2 expression, accumulation of extracellular HA and its assembly into pericellular matrices. Inhibition of HA synthesis by 4-methylumbelliferone (4MU), removal of the HA coat by hyaluronidase (HYAL) or HAS2 knockdown led to abrogation of SMC gene expression. HA activates ERK1/2 and suppresses EGFR signaling pathways via its principle receptor, CD44. EGFR inactivation coincided with increased binding to CD44, which was further augmented by addition of high molecular weight (HMW)-HA either exogenously or via HAS2 overexpression through adenoviral gene transfer. HMW-HA-stimulated esSMCs displayed a functional role in vascular tissue engineering ex vivo, vasculogenesis in a matrigel plug model and SMC accumulation in neointimal lesions of vein grafts in mice. These findings demonstrate that HAS2-induced HA synthesis and organization drives ESC-SMC differentiation. Thus, remodeling of the HA microenvironment is a critical step in directing stem cell differentiation toward a vascular lineage, highlighting HA as a potential target for treatment of vascular diseases. Stem Cells 2016 Embryonic stem cell-derived smooth muscle cells (esSMCs) maintained in high molecular weight HA (HMW-HA) were mixed with embryonic stem cell-derived endothelial cells (esECs) and then added to a Matrigel plug before subcutaneous injection into a mouse. After 1 week the Matrigel plug was removed and sections were stained for endothelial marker CD31 (green). esSMCs were labelled with Vybrant (red) before implantation in order to be distinguished from esECs. Vybrant stained esSMCs interacted with esECs to form dense and robust tube-like structures. DAPI (blue). bar, 100 μm.

Original languageEnglish
JournalStem cells (Dayton, Ohio)
Early online date4 Mar 2016
Publication statusE-pub ahead of print - 4 Mar 2016


  • Hyaluronan
  • Neointima
  • Smooth muscle cells
  • Stem cells
  • Vasculogenesis


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