Key result
New protocol enables effective single-nuclei isolation from fragile adipose tissue for downstream RNA sequencing.
Why the study?
Unbiased decomposition of adipose tissues into cellular subtypes using single-cell genomics has been challenging due to the fragile nature of lipid-filled adipocytes.
Provides a methodological protocol to overcome the fragility of lipid-filled adipocytes, enabling single-nucleus RNA sequencing to study cellular heterogeneity in adipose tissue.
Heterogeneity of thermogenic adipocytes may refine metabolic targets; leaves open distinct roles in cardiometabolic risk.
Brown and beige fat are specialized adipose tissues that dissipate energy for thermogenesis by UCP1 (Uncoupling Protein-1)-dependent and independent pathways. Until recently, thermogenic adipocytes were considered a homogeneous population. However, recent studies have indicated that there are multiple subtypes or subpopulations that are distinct in developmental origin, substrate use, and transcriptome. Despite advances in single-cell genomics, unbiased decomposition of adipose tissues into cellular subtypes has been challenging because of the fragile nature of lipid-filled adipocytes. The protocol presented was developed to circumvent these obstacles by effective isolation of single nuclei from adipose tissue for downstream applications, including RNA sequencing. Cellular heterogeneity can then be analyzed by RNA sequencing and bioinformatic analyses.
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Benitez et al. (2020) studied this question. Protocol for isolation of single nuclei from adipose tissue was evaluated. A protocol was developed for the effective isolation of single nuclei from fragile, lipid-filled adipose tissue to enable downstream single-cell genomic applications such as RNA sequencing.
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