Adult zebrafish retinal cell culture

Scope of the method

The Method relates to
  • Animal health
The Method is situated in
  • Basic Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Animal derived cells / tissues / organs
Species from which cells/tissues/organs are derived
Zebrafish (Danio rerio)
Type of cells/tissues/organs


Method keywords
  • retina
  • cell culture
  • neurite outgrowth
  • microfluidics
  • Neurons
  • zebrafish
  • transgenic lines
  • compartment-specific treatment
  • network formation
Scientific area keywords
  • axonal regeneration
  • bioenergetics
  • intraneuronal remodeling
  • dendritic remodeling
  • bio-imaging
Method description

Since adult zebrafish retinal ganglion cells (RGCs) can fully regenerate upon axonal injury, these neurons form the ideal subject to study what is driving the recovery process. The use of an adult zebrafish retinal cell culture in a microfluidic set-up enables to create a neuronal network, mimicking the normal neuronal environment. Additionally, it allows to visualize/interfere with specific intraneuronal compartments, providing a clear advantage compared to in vivo models. Overall, this state-of-the-art setup facilitates the study of processes associated with spontaneous regeneration at a single-RGC level, and high-throughput in vitro screening of potential pro-regenerative/neuroprotective therapeutic targets on a selected set of neurons. The protocol includes: retinal dissection and dissociation, cell culturing, immunostainings, and (confocal) microscopy.

Lab equipment
  • Microfluidic neuronal culturing devices ;
  • Inverted (confocal) microscope ;
  • Horizontal laminar flow ;
  • Sterile biological safety cabinet ;
  • Specific cell incubator ;
  • Dissection microscope.
Method status
  • Still in development

Pros, cons & Future potential


This retinal cell culture provides an animal saving strategy, where in addition, a neuronal network is created between two populations of neurons, mimicking the in vivo neuronal environment. Lastly, this microfluidic device enables easy and high-throughput screening and facilitated directed compound administration.


As we are the first to try this new technique of culturing adult zebrafish retinal neurons (in a microfluidic setup) a lot of optimization and validation steps are required along the process.


Once this protocol is optimized, we will build our own designed microfluidic setup, to fit all the aspects of our research question.

Future & Other applications

This set-up allows the incorporation of a neuronal population of choice in the neuronal network, thereby providing primary information about the effect of selected compounds on neuronal survival/regeneration in an in vivo simulated environment.

References, associated documents and other information


V. Grozdanov, A. Muller, V. Sengottuvel, M. Leibinger, D. Fischer A method for preparing primary retinal cell cultures for evaluating the neuroprotective and neuritogenic effect of factors on axotomized mature CNS neurons Curr. Protoc. Neurosci., Chapter 3 (2010) Unit 3.22

Other remarks

Method not published yet

Contact person

Annelies Van Dyck


Katholieke Universiteit Leuven (KUL)
Flemish Region