Embryonic stem cells isolated from Atlantic cod (Gadus morhua) and the developmental expression of a stage-specific transcription factor ac-Pou2
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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.
KeywordsAtlantic cod Embryonic stem cells ac-Pou2 Stage-specific genetic marker
The cooperation with Marine Harvest Cod and Sagafjord Seafarm is highly appreciated. The always helpful Julie Skadal at the hatching facility, Ilab, Bergen and the technical assistance of Synnøve Winterthun, Natalia Larsen and Hui-Shan Tung, NIFES, are highly valued. Special thanks to Pål A. Olsvik, NIFES, for the use of cod-specific HSP90β and HSC70 primers. Research on freshly prepared fish ESC and yolk sac larvae is allowed according to The National Animal Research Authority in Norway. This paper was financed by NIFES and Norwegian Research Council (project number: 173534/I30).
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