Document Detail


Bovine blastocyst-derived trophectoderm and endoderm cell cultures: interferon tau and transferrin expression as respective in vitro markers.
MedLine Citation:
PMID:  10642558     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Continuous cultures of bovine trophectoderm (CT-1 and CT-5) and bovine endoderm (CE-1 and CE-2) were initiated and maintained on STO feeder cells. CT-1 and CT-5 were derived from the culture of intact, 10- to 11-day in vitro-produced blastocysts. CE-1 and CE-2 were derived from the culture of immunodissected inner cell masses of 7- to 8-day in vitro-produced blastocysts. The cultures were routinely passaged by physical dissociation. Although morphologically distinct, the trophectoderm and endoderm both grew as cell sheets of polarized epithelium (dome formations) composed of approximately cuboidal cells. Both cell types, particularly the endoderm, grew on top of the feeder cells for the most part. Trophectoderm cultures grew faster, relative to endoderm, in large, rapidly extending colonies of initially flat cells with little or no visible lipid. The endoderm, in contrast, grew more slowly as tightly knit colonies with numerous lipid vacuoles in the cells at the colony centers. Ultrastructure analysis revealed that both cell types were connected by desmosomes and tight junctional areas, although these were more extensive in the trophectoderm. Endoderm was particularly rich in rough endoplasmic reticulum and Golgi apparatus indicative of cells engaged in high protein production and secretion. Interferon tau expression was specific to trophectoderm cultures, as demonstrated by reverse transcription-polymerase chain reaction, Western blot, and antiviral activity; and this property may act as a marker for this cell type. Serum protein production specific to endoderm cultures was demonstrated by Western blot; this attribute may be a useful marker for this cell type. This simple coculture method for the in vitro propagation of bovine trophectoderm and endoderm provides a system for assessing their biology in vitro.
Authors:
N C Talbot; T J Caperna; J L Edwards; W Garrett; K D Wells; A D Ealy
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biology of reproduction     Volume:  62     ISSN:  0006-3363     ISO Abbreviation:  Biol. Reprod.     Publication Date:  2000 Feb 
Date Detail:
Created Date:  2000-02-25     Completed Date:  2000-02-25     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  0207224     Medline TA:  Biol Reprod     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  235-47     Citation Subset:  IM    
Affiliation:
USDA, ARS, LPSI, Gene Evaluation and Mapping Laboratory, and Growth Biology Laboratory, Beltsville Agricultural Research Center, Beltsville, Maryland 20705, USA. ntalbot@lpsi.barc.usda.gov
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MeSH Terms
Descriptor/Qualifier:
Animals
Antiviral Agents / pharmacology
Biological Markers
Blastocyst / cytology,  physiology*,  ultrastructure
Cattle
Cells, Cultured
Culture Media, Conditioned
Cytogenetics
DNA / biosynthesis,  genetics
Endoderm / cytology,  physiology*,  ultrastructure
Gene Expression Regulation / genetics
Immunoblotting
Interferon Type I / biosynthesis*,  genetics,  pharmacology
Karyotyping
Microscopy, Electron
Pregnancy Proteins / biosynthesis*,  genetics,  pharmacology
Reverse Transcriptase Polymerase Chain Reaction
Transferrin / biosynthesis*,  genetics
Viruses / drug effects
Chemical
Reg. No./Substance:
0/Antiviral Agents; 0/Biological Markers; 0/Culture Media, Conditioned; 0/Interferon Type I; 0/Pregnancy Proteins; 0/trophoblastin; 11096-37-0/Transferrin; 9007-49-2/DNA

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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