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

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

STAR protocols (2021-11-30)
Aida Menéndez-Méndez, Lucía Paniagua-Herranz, Luis A Olivos-Oré, Rosa Gómez-Villafuertes, Raquel Pérez-Sen, Esmerilda G Delicado, Antonio R Artalejo, Felipe Ortega
摘要

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).

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Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
牛血清白蛋白 来源于牛血清, lyophilized powder, BioReagent, suitable for cell culture
Sigma-Aldrich
3-(2-氨乙酸)吲哚 盐酸盐, powder
Sigma-Aldrich
胶质纤维酸性蛋白(GFAP)单克隆抗体 小鼠抗, clone G-A-5, ascites fluid
Sigma-Aldrich
单克隆抗 β-微管蛋白 III 小鼠抗, clone SDL.3D10, ascites fluid
Sigma-Aldrich
5′-三磷酸鸟苷三钠三磷酸鸟苷三钠 锂盐, ~95% (HPLC), powder