Work

Protocol for isolating nuclei from human stem cell-derived grafts for single-nucleus RNA sequencing

Year: 2025

Type: article

Abstract: Single-nucleus RNA sequencing enables high-resolution transcriptomic profiling of brain tissue, facilitating analysis of cell identity in models of neurodegeneration and repair. Here, we present a protocol for isolating nuclei from long-term human stem cell-derived grafts in the rat brain. We describe steps for vibratome sectioning, graft dissection, nuclear extraction, and fluorescence-activated sorting. This workflow supports analysis of human neurons embedded within host tissue or sensitive to dissociation, enabling assessment of graft composition, integration, and neuronal identity in the living brain. For complete details on the use and execution of this protocol, please refer to Fiorenzano et al. 1

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104197

Authors: Edoardo Sozzi, Petter Storm, Alessandro Fiorenzano

Institutions: Lund University, University of Naples Federico II

Language: English

Topic: Single-cell and spatial transcriptomics

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,550

Funders: Parkinson's Foundation, Vetenskapsrådet, Kungliga Fysiografiska Sällskapet i Lund, Parkinsonfonden