用于组学研究的多种脑类细胞的核分离。
Nuclei isolation of multiple brain cell types for omics interrogation.
机构信息
Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA.
Department of Brain Sciences, Imperial College London, White City Campus, London, UK.
出版信息
Nat Protoc. 2021 Mar;16(3):1629-1646. doi: 10.1038/s41596-020-00472-3. Epub 2021 Jan 25.
We present a nuclei isolation protocol for genomic and epigenomic interrogation of multiple cell type populations in the human and rodent brain. The nuclei isolation protocol allows cell type-specific profiling of neurons, microglia, oligodendrocytes, and astrocytes, being compatible with fresh and frozen samples obtained from either resected or postmortem brain tissue. This 2-day procedure consists of tissue homogenization with fixation, nuclei extraction, and antibody staining followed by fluorescence-activated nuclei sorting (FANS) and does not require specialized skillsets. Cell type-specific nuclei populations can be used for downstream omic-scale sequencing applications with an emphasis on epigenomic interrogation such as histone modifications, transcription factor binding, chromatin accessibility, and chromosome architecture. The nuclei isolation protocol enables translational examination of archived healthy and diseased brain specimens through utilization of existing medical biorepositories.
我们提出了一种核分离方案，用于对人类和啮齿动物大脑中的多种细胞类型群体进行基因组和表观基因组研究。该核分离方案允许对神经元、小胶质细胞、少突胶质细胞和星形胶质细胞进行细胞类型特异性分析，与从切除或死后脑组织中获得的新鲜和冷冻样本兼容。这个为期两天的程序包括用固定剂进行组织匀浆、核提取和抗体染色，然后进行荧光激活核分选（FANS），不需要专门的技能。细胞类型特异性核群体可用于下游的组学测序应用，重点是表观基因组研究，如组蛋白修饰、转录因子结合、染色质可及性和染色体结构。核分离方案通过利用现有的医学生物库，实现了对存档的健康和患病脑组织标本的转化研究。
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