Embryonic Stem Cell Protocols pp 59-79

Part of the Methods in Molecular Biology book series (MIMB, volume 329)

Derivation, Maintenance, and Induction of the Differentiation In Vitro of Equine Embryonic Stem Cells

  • Shigeo Saito
  • Ken Sawai
  • Arika Minamihashi
  • Hideyo Ugai
  • Takehide Murata
  • Kazunari K. Yokoyama

Abstract

We describe here the isolation and maintenance of pluripotent embryonic stem (ES) cells from equine blastocysts that have been frozen and thawed. Equine ES cells appear to main-tain a normal diploid karyotype in culture. These cells express markers that are characteris-tic of mouse ES cells, namely, alkaline phosphatase, stage-specific-embryonic antigen 1, STAT3, and Oct4. We also describe protocols for the induction of differentiation in vitro to neural precursor cells in the presence of basic fibroblast growth factor (bFGF), epidermal growth factor, and platelet-derived growth factor and to hematopoietic and endothelial cell lineages in the presence of bFGF, stem cell factor, and oncostatin M. Equine ES cells pro-vide a powerful tool for gene targeting and the generation of transgenic clonal offspring.

Key Words

Alkaline phosphatase bFGF CD34 CD35 CD45 derivation EGF endothelial differentiation equine blastocysts equine ES cells Flk-1 GATA 4 GFAP hematopoietic differentiation LIF maintenance Nestin neural differentiation Oct4 OSM PDGF pluripotency SCF Sry-1 SSEA-1 STAT3 β -tubulin III. 

Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Shigeo Saito
    • 1
  • Ken Sawai
    • 2
  • Arika Minamihashi
    • 3
  • Hideyo Ugai
    • 4
  • Takehide Murata
    • 5
  • Kazunari K. Yokoyama
    • 6
  1. 1.Saito Laboratory of Cell TechnologyKataoka, Yaita, TechigiJapan
  2. 2.Hokkaido Animal ResearchShintoku, HokkaidoJapan
  3. 3.Hokkaido Animal ResearchShintoku, HokkaidoJapan
  4. 4.Gene Engineering Division, BioResourceRIKEN (The Institute of Physical and Chemical Research)IbarakiJapan
  5. 5.Gene Engineering Division, BioResource CentreRIKEN (The Institute of Physical and Chemical Research)IbarakiJapan
  6. 6.Gene Engineering Division, BioResourceRIKEN (The Institute of Physical and Chemical Research)IbarakiJapan

Personalised recommendations