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Updated: Jul 2, 2025
Isolation of Adipose Tissue Nuclei for Single-Cell Genomic Applications
Published on: June 12, 2020
Protocol for flow cytometry-assisted single-nucleus RNA sequencing of human and mouse adipose tissue with sample
Anton Gulko1, Adam Essene1, Dylan Matthew Belmont-Rausch2
1Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA; Boston Nutrition and Obesity Research Center/Boston Area Diabetes and Endocrinology Research Center Functional Genomics and Bioinformatics Core, Boston, MA, USA.
Abstract:
Adipocyte size and fragility and commercial kit costs impose significant limitations on single-cell RNA sequencing of adipose tissue. Accordingly, we developed a workflow to isolate and sample-barcode nuclei from individual adipose tissue samples, integrating flow cytometry for quality control, counting, and precise nuclei pooling for direct loading onto the popular 10× Chromium controller. This approach can eliminate batch confounding, and significantly reduces poor-quality nuclei, ambient RNA contamination, and droplet loading-associated reagent waste, resulting in pronounced improvements in information content and cost efficiency.
