Department of Biochemistry and Molecular Biology, Faculty of Veterinary, Universidad Complutense de Madrid (UCM), 28040 Madrid, Spain.
Instituto Universitario de Investigación en Neuroquímica (IUIN), 28029 Madrid, Spain.
STAR Protoc. 2021 Nov 18;2(4):100964. doi: 10.1016/j.xpro.2021.100964. eCollection 2021 Dec 17.
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 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).
对出生后小脑进行低密度细胞培养,结合活体成像和单细胞跟踪,可监测出生后小脑神经干细胞(NSC)及其后代的行为。培养的小脑 NSC 保持其神经发生特性,以与由三个出生后小脑神经发生龛产生的神经元后代相同的相对比例产生后代。本方案通过成像后免疫细胞化学和膜片钳记录来描述后代的性质。有关使用和执行此方案的完整详细信息,请参见 Paniagua-Herranz 等人(2020b)。