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
-
- Zebrafish/embryology*
- Macrophages/cytology
- Flow Cytometry
- Purkinje Cells/cytology
- Electroporation
- Motor Neurons/cytology
- Subcellular Fractions/metabolism
- Animals
- Primary Cell Culture/methods*
- Cells, Cultured
- Imaging, Three-Dimensional*
- Staining and Labeling
- Cell Shape
- Neuroglia/cytology
- Transgenes
- Transfection/methods*
- Embryo, Nonmammalian/cytology*
- Embryo, Nonmammalian/metabolism
- PubMed
- 28802829 Full text @ Dev. Biol.
Citation
Sassen, W.A., Lehne, F., Russo, G., Wargenau, S., Dübel, S., Köster, R.W. (2017) Embryonic zebrafish primary cell culture for transfection and live cellular and subcellular imaging. Developmental Biology. 430(1):18-31.
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping