Combining low-density cell culture, single-cell tracking, and patch-clamp to monitor the behavior of postnatal murine cerebellar neural stem cells

• Published in: STAR protocols 2021-12, Vol.2 (4), p.100964, Article 100964
• Main Authors: Menéndez-Méndez, Aida, Paniagua-Herranz, Lucía, Olivos-Oré, Luis A., Gómez-Villafuertes, Rosa, Pérez-Sen, Raquel, Delicado, Esmerilda G., Artalejo, Antonio R., Ortega, Felipe
• Format: Article
• Language: English
• Subjects: Animals; Cell Biology; Cell culture; Cells, Cultured; Cerebellum - cytology; Cytological Techniques - methods; Developmental biology; Female; Male; Mice; Mice, Inbred C57BL; Microscopy; Neural Stem Cells - cytology; Neuroscience; Protocol; Single Cell; Stem Cells
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2021.100964

Low-density cell culture of the postnatal cerebellum, combined with live imaging and single-cell tracking, allows the behavior of postnatal cerebellar neural stem cells (NSCs) and their progeny to be monitored. Cultured cerebellar NSCs maintain their neurogenic nature giving rise, in the same relative proportions that exist in vivo, to the neuronal progeny generated by the three postnatal cerebellar neurogenic niches. This protocol describes the identification of the nature of the progeny through both post-imaging immunocytochemistry and patch-clamp recordings. For complete details on the use and execution of this protocol, please refer to Paniagua-Herranz et al. (2020b). [Display omitted] •Protocol to culture postnatal mouse cerebellar neural stem cells (NSCs) in low density•The behavior of isolated postnatal cerebellar NSC can be monitored at single-cell level•Protocol for simultaneous monitoring of the three postnatal cerebellar neurogenic niches•Procedures of live imaging and single cell tracking for lineage tracing Low-density cell culture of the postnatal cerebellum, combined with live imaging and single-cell tracking, allows the behavior of postnatal cerebellar neural stem cells (NSCs) and their progeny to be monitored. Cultured cerebellar NSCs maintain their neurogenic nature giving rise, in the same relative proportions that exist in vivo, to the neuronal progeny generated by the three postnatal cerebellar neurogenic niches. This protocol describes the identification of the nature of the progeny through both post-imaging immunocytochemistry and patch-clamp recordings.