Nuclei isolation from surgically resected human hippocampus

STAR Protoc. 2021 Sep 16;2(4):100844. doi: 10.1016/j.xpro.2021.100844. eCollection 2021 Dec 17.

Abstract

Single-nucleus RNA sequencing (snRNA-seq), where nuclear transcriptomes are a proxy to cellular transcriptomes, has been used to profile human brain. snRNA-seq is sensitive to tissue processing, tissue quality, postmortem interval time, and cellular debris. This protocol outlines steps for the isolation of high-quality nuclei from surgically resected human brain tissue followed by a sucrose gradient yielding neuronal and non-neuronal nuclei enabling unbiased analysis of various cell types. For complete details on the use and execution of this protocol, please refer to Ayhan et al. (2021).

Keywords: Cell isolation; Cell separation/fractionation; Genomics; Microbiology; Neuroscience; RNAseq; Single Cell.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus* / metabolism
  • Hippocampus* / surgery
  • Humans
  • RNA, Small Nuclear / metabolism
  • Sequence Analysis, RNA / methods
  • Solitary Nucleus

Substances

  • RNA, Small Nuclear