Fish embryo cell cultures for derivation of stem cells and transgenic chimeras.
It has been demonstrated in mammalian systems that techniques using embryonal stem cells provide advantages over conventional injection of DNA into embryos for generation of transgenic animals. We employed cell culture approaches in an attempt to develop this technology for fish transgenesis. Using a trout embryo-derived mitogenic preparation in a specialized culture medium, we initiated replication of zebrafish blastula-derived cell cultures and expressed marker genes introduced into the cells by plasmid transfection. Reintroduction of cells from the cultures into blastula-stage embryos indicated that the cultured cells survived and may contribute to the developing organism.
Collodi
P
P
Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331.
Kamei
Y
Y
Sharps
A
A
Weber
D
D
Barnes
D
D
D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology* / Q000235:genetics; D001755:Blastocyst / Q000166:cytology / Q000201:enzymology; D002478:Cells, Cultured; D002678:Chimera; D016895:Culture Media, Serum-Free; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000201:enzymology; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression; D006238:Haploidy; D007621:Karyotyping; D008297:Male; D010957:Plasmids / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000502:physiology; D033581:Stem Cell Transplantation; D014162:Transfection; D014337:Trout / Q000196:embryology* / Q000235:genetics; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics; D001616:beta-Galactosidase / Q000096:biosynthesis
D016895:Culture Media, Serum-Free; D001616:beta-Galactosidase