TY - JOUR
T1 - Establishment of cell lines from individual zebrafish embryos
AU - Geyer, Nathalie
AU - Kaminsky, Sabrina
AU - Confino, Shir
AU - Livne, Zohar Ben Moshe
AU - Gothilf, Yoav
AU - Foulkes, Nicholas S.
AU - Vallone, Daniela
N1 - Publisher Copyright:
© The Author(s) 2023.
PY - 2023/10
Y1 - 2023/10
N2 - With the increasing use of fish as model species for research, cell cultures derived from caudal fin explants as well as pre-hatching stage embryos have provided powerful in vitro tools that can complement or serve as an ethically more acceptable alternative to live animal experiments. The widely-used protocols to establish these lines require, as a starting point, homogeneous pools of embryos or viable adult fish which are large enough for collecting sufficient fin tissue. This excludes the use of fish lines with adverse phenotypes or lines that exhibit mortality at early developmental stages and so can only be propagated as heterozygotes. Specifically, when no visually overt mutant phenotype is detectable for identifying homozygous mutants at early embryonic stages, it is then impossible to sort pools of embryos with the same genotypes to generate cell lines from the progeny of a heterozygote in-cross. Here, we describe a simple protocol to generate cell lines on a large scale starting from individual early embryos that can subsequently be genotyped by polymerase chain reaction. This protocol should help to establish fish cell culture models as a routine approach for the functional characterization of genetic changes in fish models such as the zebrafish. Furthermore, it should contribute to a reduction of experiments which are ethically discouraged to avoid pain and distress.
AB - With the increasing use of fish as model species for research, cell cultures derived from caudal fin explants as well as pre-hatching stage embryos have provided powerful in vitro tools that can complement or serve as an ethically more acceptable alternative to live animal experiments. The widely-used protocols to establish these lines require, as a starting point, homogeneous pools of embryos or viable adult fish which are large enough for collecting sufficient fin tissue. This excludes the use of fish lines with adverse phenotypes or lines that exhibit mortality at early developmental stages and so can only be propagated as heterozygotes. Specifically, when no visually overt mutant phenotype is detectable for identifying homozygous mutants at early embryonic stages, it is then impossible to sort pools of embryos with the same genotypes to generate cell lines from the progeny of a heterozygote in-cross. Here, we describe a simple protocol to generate cell lines on a large scale starting from individual early embryos that can subsequently be genotyped by polymerase chain reaction. This protocol should help to establish fish cell culture models as a routine approach for the functional characterization of genetic changes in fish models such as the zebrafish. Furthermore, it should contribute to a reduction of experiments which are ethically discouraged to avoid pain and distress.
KW - Fish cell culture
KW - early-stage mutant embryos
KW - zebrafish embryos
UR - http://www.scopus.com/inward/record.url?scp=85150854577&partnerID=8YFLogxK
U2 - 10.1177/00236772231157162
DO - 10.1177/00236772231157162
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
C2 - 36896487
AN - SCOPUS:85150854577
SN - 0023-6772
VL - 57
SP - 518
EP - 528
JO - Laboratory Animals
JF - Laboratory Animals
IS - 5
ER -