Zebrafish Embryo Cells Remain Pluripotent and Germ-Line Competent for Multiple Passages in Culture
- 1 February 2004
- journal article
- Published by Mary Ann Liebert Inc in Zebrafish
- Vol. 1 (1) , 21-26
- https://doi.org/10.1089/154585404774101644
Abstract
Mouse embryonic stem (ES) cell lines are routinely used to introduce targeted mutations into the genome, providing an efficient method to study gene function. Application of similar gene knockout techniques to other organisms has been unsuccessful due to the lack of germ-line competent ES cell lines from non-murine species. Previously, we reported the production of zebrafish germ-line chimeras using short-term primary embryo cell cultures. Here we demonstrate that zebrafish embryo cells, maintained for several weeks and multiple passages in culture, remain pluripotent and germ-line competent. Zebrafish germ-line chimeras were generated from passage 5 and 6 cultures initiated from blastula- and gastrula-stage embryos. In addition to the germ line, the cultured cells contributed to multiple tissues of the host embryo, including muscle, liver, gut, and fin. To facilitate the identification of germ-line chimeras, ES cells expressing the green fluorescent protein (GFP) were introduced into host embryos, and germ-line contribution was detected by the presence of GFP+ cells in the region of the gonad. The germ-line competent embryo cell cultures will be useful for the development of a gene targeting strategy that will increase the utility of the zebrafish model for studies of gene function.Keywords
This publication has 25 references indexed in Scilit:
- Target-Selected Inactivation of the Zebrafish rag1 GeneScience, 2002
- Insertional mutagenesis in zebrafish rapidly identifies genes essential for early vertebrate developmentNature Genetics, 2002
- Production of α-1,3-Galactosyltransferase Knockout Pigs by Nuclear Transfer CloningScience, 2002
- Zebrafish--the Canonical VertebrateScience, 2001
- Effective targeted gene ‘knockdown’ in zebrafishNature Genetics, 2000
- Zebrafish genetics: Mutant cornucopiaCurrent Biology, 1996
- Of Flies and FishesScience, 1994
- Altering the Genome by Homologous RecombinationScience, 1989
- Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genesNature, 1988
- Production of clones of homozygous diploid zebra fish (Brachydanio rerio)Nature, 1981