TitleSynthesis and organization of hyaluronan and versican by embryonic stem cells undergoing embryoid body differentiation.
Publication TypeJournal Article
Year of Publication2010
AuthorsShukla, S, Nair, R, Rolle, MW, Braun, KR, Chan, CK, Johnson, PY, Wight, TN, McDevitt, TC
JournalJournal of Histochemistry & Cytochemistry
Date PublishedApril 2010
KeywordsAnimals, Cell Differentiation, Cell Line, Embryonic Stem Cells, Epithelial Cells, Hyaluronic Acid, Mesenchymal Stromal Cells, Mice, Versicans

Embryonic stem cells (ESCs) provide a convenient model to probe the molecular and cellular dynamics of developmental cell morphogenesis. ESC differentiation in vitro via embryoid bodies (EBs) recapitulates many aspects of early stages of development, including the epithelial-mesenchymal transition (EMT) of pluripotent cells into more differentiated progeny. Hyaluronan and versican are important extracellular mediators of EMT processes, yet the temporal expression and spatial distribution of these extracellular matrix (ECM) molecules during EB differentiation remains undefined. Thus, the objective of this study was to evaluate the synthesis and organization of hyaluronan and versican by using murine ESCs during EB differentiation. Hyaluronan and versican (V0 and V1 isoforms), visualized by immunohistochemistry and evaluated biochemically, accumulated within EBs during the course of differentiation. Interestingly, increasing amounts of a 70-kDa proteolytic fragment of versican were also detected over time, along with ADAMTS-1 and -5 protein expression. ESCs expressed each of the hyaluronan synthases (HAS) -1, -2, and -3 and versican splice variants (V0, V1, V2, and V3) throughout EB differentiation, but HAS-2, V0, and V1 were expressed at significantly increased levels at each time point examined. Hyaluronan and versican exhibited overlapping expression patterns within EBs in regions of low cell density, and versican expression was excluded from clusters of epithelial (cytokeratin-positive) cells but was enriched within the vicinity of mesenchymal (N-cadherin-positive) cells. These results indicate that hyaluronan and versican synthesized by ESCs within EB microenvironments are associated with EMT processes and furthermore suggest that endogenously produced ECM molecules play a role in ESC differentiation. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.

Alternate JournalJ. Histochem. Cytochem.
PubMed ID20026669
PubMed Central IDPMC2842597
Grant ListR24 HL64387-06A1 / HL / NHLBI NIH HHS / United States
R21 EB007316 / EB / NIBIB NIH HHS / United States
F32 HL083593 / HL / NHLBI NIH HHS / United States