PUBLICATION

Zebrafish Embryo Cells Remain Pluripotent and Germ-Line Competent for Multiple Passages in Culture

Authors
Fan, L., Crodian, J., Liu, X., Aleström, A., Aleström, P., and Collodi, P.
ID
ZDB-PUB-040521-2
Date
2004
Source
Zebrafish   1(1): 21-26 (Journal)
Registered Authors
Aleström, Peter, Collodi, Paul, Crodian, Jennifer, Fan, Lianchun, Liu, Xiangyu
Keywords
none
MeSH Terms
none
PubMed
18248202 Full text @ Zebrafish
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.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping