ZFIN ID: ZDB-PUB-061227-17
Zebrafish embryonic stem cells
Fan, L., and Collodi, P.
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.
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.