Single-cell RNA-sequencing of ChAT-eGFP+ adipose stromal vascular cells

ChAT-eGFP mice were housed at thermoneutrality for three weeks then either kept at thermoneutrality or exposed to acute cold challenge, then inguinal adipose stromal vascular cells were harvested and scRNAseq performed on sorted ChAT-eGFP+ cells. This study represents the first single-cell transcriptomic profiling of cholinergic cells in fat and seeks to understand the regulation of these cells in response to acute cold exposure and thermogenic activation. Overall design: At five weeks of age, male ChAT-eGFP mice were housed at thermoneutrality (30C) for three weeks, then either kept at thermoneutrality or exposed to acute cold challenge (4C) for 4h. Two biological replicates per condition were used, with each biological replicate comprised of cells from four pooled mice. Inguinal subcutaneous white adipose tissue was harvested then stromal vascular cells were isolated following tissue digestion. Live, single ChAT-eGFP+ cells were FACS-sorted then ~10,000 cells per biological replicate were submitted for 10x Genomics scRNAseq (3' GEX v3).