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Human embryonic stem cells in culture possess primary cilia with hedgehog signaling machinery

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  • Enko N Kiprilov
  • Aashir Awan
  • Romain Desprat
  • Michelle Velho
  • Christian A Clement
  • Anne Grete Byskov
  • Claus Y Andersen
  • Peter Satir
  • Eric E Bouhassira
  • Søren T Christensen
  • Rhoda Elison Hirsch
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Human embryonic stem cells (hESCs) are potential therapeutic tools and models of human development. With a growing interest in primary cilia in signal transduction pathways that are crucial for embryological development and tissue differentiation and interest in mechanisms regulating human hESC differentiation, demonstrating the existence of primary cilia and the localization of signaling components in undifferentiated hESCs establishes a mechanistic basis for the regulation of hESC differentiation. Using electron microscopy (EM), immunofluorescence, and confocal microscopies, we show that primary cilia are present in three undifferentiated hESC lines. EM reveals the characteristic 9 + 0 axoneme. The number and length of cilia increase after serum starvation. Important components of the hedgehog (Hh) pathway, including smoothened, patched 1 (Ptc1), and Gli1 and 2, are present in the cilia. Stimulation of the pathway results in the concerted movement of Ptc1 out of, and smoothened into, the primary cilium as well as up-regulation of GLI1 and PTC1. These findings show that hESCs contain primary cilia associated with working Hh machinery.

Original languageEnglish
JournalJournal of Cell Biology
Volume180
Issue number5
Pages (from-to)897-904
Number of pages8
ISSN0021-9525
DOIs
Publication statusPublished - 10 Mar 2008

    Research areas

  • Axoneme/genetics, Cell Differentiation/genetics, Cell Line, Cell Lineage/genetics, Cilia/genetics, Culture Media, Serum-Free/pharmacology, Embryonic Stem Cells/metabolism, Female, Fluorescent Antibody Technique, Hedgehog Proteins/genetics, Humans, Kruppel-Like Transcription Factors/genetics, Male, Microscopy, Confocal, Microscopy, Electron, Transmission, Nuclear Proteins/genetics, Patched Receptors, Patched-1 Receptor, Receptors, Cell Surface/genetics, Signal Transduction/genetics, Transcription Factors/genetics, Zinc Finger Protein GLI1, Zinc Finger Protein Gli2

ID: 54910486