Optimized Primary Culture of Neuronal Populations for Subcellular Omics Applications

Primary cell culturePrimary cell cultures is an invaluable method frequently used to overcome challenges associated with in vivoIn vivo experiments. In zebrafishZebrafish research, in vivo live imagingImaging experiments are routine owing to the high optical transparency of embryos, and, as a result, primary cell culturePrimary cell cultures has been less utilized. However, the approach still boasts powerful advantages, emphasizing the importance of sophisticated zebrafishZebrafish cell culture protocols. Here, we present an enhanced protocol for the generation of primary cell culturesPrimary cell cultures by dissociation of 24 hpf zebrafishZebrafish embryos. We include a novel cell culture medium recipe specifically favoring neuronal growth and survival, enabling relatively long-term culture. We outline primary zebrafishZebrafish neuronal culture on glass coverslips, as well as in transwell insertsTranswell inserts which allow isolation of neurite tissue for experiments such as investigating subcellular transcriptomes.