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Manipulating the Avian Epiblast and Epiblast-Derived Stem Cells

  • Cantas Alev
  • Mikiharu Nakano
  • Yuping Wu
  • Hiroyuki Horiuchi
  • Guojun Sheng
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1074)

Abstract

Compared to eutherian mammals, birds retain a primitive form of epiblast development. Molecular studies of the avian epiblast can provide valuable insight for mammalian epiblast research. Here, we introduce several basic techniques in handling epiblast-stage embryos of the chick, the major model organism for avian developmental biology studies. We describe how to collect embryos for RNA extraction and gene expression analysis, to set up ex ovo New culture for overexpression, bead graft and small molecule-based inhibitor studies, and to carry out whole-mount RNA in situ hybridization analysis. We introduce a novel and simple method for molecular perturbation of the epiblast differentiation in ovo. We also describe how to perform primary chicken epiblast cell culture, to establish stable epiblast stem cell (Epi-SC) lines, and to assay for pluripotency in primary epiblast cells and Epi-SCs.

Key words

Avian Chicken Epiblast Stem cells Pluripotency New culture Bead graft In situ hybridization Chicken ES cells Epiblast stem cells 

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Cantas Alev
    • 1
  • Mikiharu Nakano
    • 2
  • Yuping Wu
    • 3
  • Hiroyuki Horiuchi
    • 2
  • Guojun Sheng
    • 3
  1. 1.Laboratory for Early EmbryogenesisRIKEN Center for Developmental BiologyChuo-Ku, KobeJapan
  2. 2.Laboratory for Immunobiology, Department of Molecular and Applied BioscienceHiroshima UniversityHigashi-HiroshimaJapan
  3. 3.Laboratory for Early EmbryogenesisRIKEN Center for Developmental BiologyChuo-Ku, KobeJapan

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