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Isolation of Nuclei from Flash-Frozen Liver Tissue for Single-Cell Multiomics
Published on: December 9, 2022
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Single-nucleus RNA-seq2 reveals functional crosstalk between liver zonation and ploidy
M L Richter1, I K Deligiannis1, K Yin1,2
1Helmholtz Pioneer Campus (HPC), Helmholtz Zentrum München, Neuherberg, Germany.
Nature Communications
|July 13, 2021
Summary
Single-nucleus RNA-seq2 analyzes liver cell heterogeneity in frozen tissues. Ploidy states in hepatocytes correlate with metabolic potential and lobule position, revealing gene dosage and spatial distribution crosstalk.
Area of Science:
- Genomics
- Cell Biology
- Hepatology
Background:
- Single-cell RNA sequencing (scRNA-seq) is crucial for understanding cellular heterogeneity in disease.
- Analyzing frozen tissues presents challenges for comprehensive cellular transcriptome analysis.
Purpose of the Study:
- To develop a single-nucleus RNA-seq2 method for analyzing frozen liver tissues.
- To characterize hepatocyte heterogeneity, focusing on ploidy and spatial distribution.
Main Methods:
- Development of a single-nucleus RNA-seq2 protocol for frozen liver samples.
- Comprehensive analysis of the nuclear transcriptome across diverse liver cell types.
- Characterization of transcriptional profiles of hepatocytes based on ploidy levels.
Main Results:
- The single-nucleus RNA-seq2 method successfully dissected all liver cell types from frozen tissues.
- Hepatocyte ploidy states were associated with distinct metabolic potentials.
- Gene expression in tetraploid hepatocytes was influenced by their position within the hepatic lobule.
Conclusions:
- Single-nucleus RNA-seq2 is effective for studying cellular heterogeneity in frozen liver tissues.
- Hepatocyte ploidy and spatial location are key determinants of metabolic function and gene expression.
- A crosstalk exists between gene dosage and spatial distribution in hepatocytes.
