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

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

By: Shigeo Saito², Ken Sawai³, Arika Minamihashi⁴, Hideyo Ugai⁵, Takehide Murata⁶, Kazunari K. Yokoyama⁷

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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.

Affiliation(s): (2) Saito Laboratory of Cell Technology, Kataoka, Yaita, Techigi, Japan
(3) Hokkaido Animal Research, Shintoku, Hokkaido, Japan
(4) Hokkaido Animal Research, Shintoku, Hokkaido, Japan
(5) Gene Engineering Division, BioResource, RIKEN (The Institute of Physical and Chemical Research), Ibaraki, Japan
(6) Gene Engineering Division, BioResource Centre, RIKEN (The Institute of Physical and Chemical Research), Ibaraki, Japan
(7) Gene Engineering Division, BioResource, RIKEN (The Institute of Physical and Chemical Research), Ibaraki, Japan

Book Title: Embryonic Stem Cell Protocols: Volume 1: Isolation and Characterization

Series: Methods in Molecular Biology | Volume: 329 | Pub. Date: Feb-15-2006 | Page Range: 59-79 | DOI: 10.1385/1-59745-037-5:59

Subject: Cell Biology

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.

References

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