Combining low-density cell culture, single-cell tracking, and patch-clamp to monitor the behavior of postnatal murine cerebellar neural stem cells
| Autor: | Felipe Ortega, Antonio R. Artalejo, Rosa Gómez-Villafuertes, Raquel Pérez-Sen, Luis Alcides Olivos-Oré, Esmerilda G. Delicado, Aida Menéndez-Méndez, Lucía Paniagua-Herranz |
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| Rok vydání: | 2021 |
| Předmět: |
Male
Cerebellum Science (General) Immunocytochemistry Cytological Techniques Single Cell Biology General Biochemistry Genetics and Molecular Biology Q1-390 Mice Neural Stem Cells Live cell imaging Developmental biology medicine Protocol Animals Patch clamp Cells Cultured Microscopy General Immunology and Microbiology General Neuroscience Stem Cells Cell Biology Neural stem cell Mice Inbred C57BL medicine.anatomical_structure nervous system Cell culture Female Stem cell Neuroscience |
| Zdroj: | STAR Protocols STAR Protocols, Vol 2, Iss 4, Pp 100964-(2021) |
| ISSN: | 2666-1667 |
| Popis: | Summary 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). Graphical abstract Highlights • 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. |
| Databáze: | OpenAIRE |
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