Document Detail

Establishment of goat embryonic stem cells from in vivo produced blastocyst-stage embryos.
MedLine Citation:
PMID:  21387453     Owner:  NLM     Status:  MEDLINE    
Embryonic stem (ES) cells with the capacity for germ line transmission have only been verified in mouse and rat. Methods for derivation, propagation, and differentiation of ES cells from domestic animals have not been fully established. Here, we describe derivation of ES cells from goat embryos. In vivo-derived embryos were cultured on goat fetal fibroblast feeders. Embryos either attached to the feeder layer or remained floating and expanded in culture. Embryos that attached showed a prominent inner cell mass (ICM) and those that remained floating formed structures resembling ICM disks surrounded by trophectodermal cells. ICM cells and embryonic disks were isolated mechanically, cultured on feeder cells in the presence of hLIF, and outgrown into ES-like colonies. Two cell lines were cultured for 25 passages and stained positive for alkaline phosphatase, POU5F1, NANOG, SOX2, SSEA-1, and SSEA-4. Embryoid bodies formed in suspension culture without hLIF. One cell line was cultured for 2 years (over 120 passages). This cell line differentiated in vitro into epithelia and neuronal cells, and could be stably transfected and selected for expression of a fluorescent marker. When cells were injected into SCID mice, teratomas were identified 5-6 weeks after transplantation. Expression of known ES cell markers, maintenance in vitro for 2 years in an undifferentiated state, differentiation in vitro, and formation of teratomas in immunodeficient mice provide evidence that the established cell line represents goat ES cells. This also is the first report of teratoma formation from large animal ES cells.
E Behboodi; A Bondareva; I Begin; K Rao; N Neveu; J T Pierson; C Wylie; F D Piero; Y J Huang; W Zeng; V Tanco; H Baldassarre; C N Karatzas; I Dobrinski
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2011-03-08
Journal Detail:
Title:  Molecular reproduction and development     Volume:  78     ISSN:  1098-2795     ISO Abbreviation:  Mol. Reprod. Dev.     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-03-22     Completed Date:  2011-07-15     Revised Date:  2013-08-26    
Medline Journal Info:
Nlm Unique ID:  8903333     Medline TA:  Mol Reprod Dev     Country:  United States    
Other Details:
Languages:  eng     Pagination:  202-11     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 Wiley-Liss, Inc.
Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta, Canada.
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MeSH Terms
Cell Culture Techniques
Cell Differentiation / physiology
Cell Separation / methods*
Embryo, Mammalian / cytology*
Embryonic Stem Cells / cytology*
Goats / embryology*
Mice, SCID
Teratoma / etiology,  pathology
Grant Support
2 R01 RR17359-06/RR/NCRR NIH HHS; 2 R42HD044780-02/HD/NICHD NIH HHS

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