Embryonic stem cells isolated from Atlantic cod (Gadus morhua) and the developmental expression of a stage-specific transcription factor ac-Pou2.
The establishment of embryonic stem cell cultures and the identification of molecular markers for undifferentiated embryonic stem cells (ESC) as well as differentiated cells types will open new opportunities in the study of developmental biology and for developing embryonic in vitro models of the ecologically and economically important fish specie Atlantic cod (Gadus morhua). We report here that cod blastula cells express a Class V POU gene known to be highly expressed in embryonic cell populations of vertebrates. The cod transcript, designated Atlantic cod-Pou2 (ac-Pou2), can be used as a genetic marker for cod blastula cells in vivo and in vitro. Using a quantitative real-time PCR approach, we found that the ac-Pou2 transcript was downregulated before the egg reached the stage of gastrulation, the starting point of extensive cell differentiation. We also demonstrate the culturing of ESC isolated from cod blastula stage eggs. The cod ESC exhibited in vitro characteristics of pluripotency described for both mammalian ESC and fish ES-like cells (medaka, zebrafish, seabream, sea perch and rainbow trout). Cod ESC in culture expressed ac-pou2, differentiated spontaneously and had the ability to form embryoid bodies following retinoic acid treatment. The ESC could also be directed to differentiate.
Holen
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E
National Institute of Nutrition and Seafood Research (NIFES), Post Box 2029 Nordnes, 5817, Bergen, Norway. eho@nifes.no
Kausland
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A
Skjærven
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K
D000595:Amino Acid Sequence; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods / Q000662:veterinary*; D002454:Cell Differentiation / Q000502:physiology; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000378:metabolism* / Q000502:physiology; D005455:Fluorescent Antibody Technique; D048389:Gadus morhua / Q000196:embryology*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D005819:Genetic Markers / Q000235:genetics*; D008969:Molecular Sequence Data; D050809:POU Domain Factors / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D013045:Species Specificity
D017931:DNA Primers; D005819:Genetic Markers; D050809:POU Domain Factors