PUBLICATION

Embryonic zebrafish primary cell culture for transfection and live cellular and subcellular imaging

Authors
Sassen, W.A., Lehne, F., Russo, G., Wargenau, S., Dübel, S., Köster, R.W.
ID
ZDB-PUB-170815-14
Date
2017
Source
Developmental Biology   430(1): 18-31 (Journal)
Registered Authors
Köster, Reinhard W., Russo, Giulio
Keywords
live cell imaging, organelle dynamics, primary cell culture, vital dyes, zebrafish
MeSH Terms
  • Animals
  • Cell Shape
  • Cells, Cultured
  • Electroporation
  • Embryo, Nonmammalian/cytology*
  • Embryo, Nonmammalian/metabolism
  • Flow Cytometry
  • Imaging, Three-Dimensional*
  • Macrophages/cytology
  • Motor Neurons/cytology
  • Neuroglia/cytology
  • Primary Cell Culture/methods*
  • Purkinje Cells/cytology
  • Staining and Labeling
  • Subcellular Fractions/metabolism
  • Transfection/methods*
  • Transgenes
  • Zebrafish/embryology*
PubMed
28802829 Full text @ Dev. Biol.
Abstract
Although having great potential for live cell imaging to address numerous cell biological questions with high spatial and temporal resolution, primary cell cultures of zebrafish embryos are not widely used. We present an easy-to-use protocol for preparing primary cell cultures of 2 dpf zebrafish embryos allowing for live cell imaging of fully differentiated cells such as neurons and myocytes. We demonstrate that different cell types can be identified by morphology and expression of transgenic cell type-specific fluorescent reporters and that fluorescent cells can be sorted by flow cytometry to prepare an enriched culture. To facilitate subcellular imaging in live primary cells, we successfully tested a selection of fluorescent vital dyes. Most importantly, we demonstrate that zebrafish primary cells can be transfected efficiently with expression constructs allowing for visualizing subcellular structures with fluorescent marker proteins for time lapse imaging. We propose zebrafish primary cell culture as a versatile tool to address cell biological questions in combination with a powerful in vivo model.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping