PUBLICATION

Optimized Primary Culture of Neuronal Populations for Subcellular Omics Applications

Authors
Taylor, R., Houart, C.
ID
ZDB-PUB-230906-57
Date
2024
Source
Methods in molecular biology (Clifton, N.J.)   2707: 113124113-124 (Chapter)
Registered Authors
Houart, Corinne
Keywords
Cell culture, Compartmentalized culture, Embryo dissociation, RNA extraction, RNAseq, Transwell Insert, Zebrafish
MeSH Terms
  • Animals
  • Cell Culture Techniques
  • Glass
  • Neurites*
  • Neurogenesis
  • Zebrafish*
PubMed
37668908 Full text @ Meth. Mol. Biol.
Abstract
Primary cell culture is an invaluable method frequently used to overcome challenges associated with in vivo experiments. In zebrafish research, in vivo live imaging experiments are routine owing to the high optical transparency of embryos, and, as a result, primary cell culture has been less utilized. However, the approach still boasts powerful advantages, emphasizing the importance of sophisticated zebrafish cell culture protocols. Here, we present an enhanced protocol for the generation of primary cell cultures by dissociation of 24 hpf zebrafish embryos. We include a novel cell culture medium recipe specifically favoring neuronal growth and survival, enabling relatively long-term culture. We outline primary zebrafish neuronal culture on glass coverslips, as well as in transwell inserts which allow isolation of neurite tissue for experiments such as investigating subcellular transcriptomes.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping