PUBLICATION
Zebrafish embryonic stem cells
- Authors
- Fan, L., and Collodi, P.
- ID
- ZDB-PUB-061227-17
- Date
- 2006
- Source
- Methods in enzymology 418(1): 64-77 (Journal)
- Registered Authors
- Collodi, Paul, Fan, Lianchun
- Keywords
- none
- MeSH Terms
-
- Animals
- Blastula/cytology
- Blastula/physiology
- Cell Culture Techniques/methods
- Culture Media
- Embryo, Nonmammalian
- Embryonic Development/physiology
- Embryonic Stem Cells/cytology*
- Gastrula/cytology
- Gastrula/physiology
- Zebrafish/embryology*
- PubMed
- 17141029 Full text @ Methods Enzymol.
Citation
Fan, L., and Collodi, P. (2006) Zebrafish embryonic stem cells. Methods in enzymology. 418(1):64-77.
Abstract
Methods are presented for the derivation of zebrafish embryonic stem (ES) cell cultures that are initiated from blastula and gastrula stage embryos. To maintain pluripotency, the ES cells are cocultured with rainbow trout spleen cells from the RTS34st cell line. ES cells maintained for multiple passages on a feeder layer of growth-arrested RTS34st exhibit in vitro characteristics of pluripotency and produce viable germ cells following transplantation into a host embryo. The ES cells are able to undergo targeted plasmid insertion by homologous recombination, and methods are described for the introduction of a targeting vector by electroporation. Two strategies are described for the efficient isolation of homologous recombinants using a visual marker screen and positive-negative selection.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping