Illumina NovaSeq 6000 paired end sequencing; scRNA-seq of stromal vascular cells from 48 week (aged) mouse subcutaneous adipose tissue compared to control 10 week (young) mouse SVF
scRNA-seq of stromal vascular cells from 48 week (aged) mouse subcutaneous adipose tissue compared to control 10 week (young) mouse SVF
Inguinal white adipose tissue (iWAT) was dissected from mice of varying ages. iWAT served as a representative subcutaneous adipose tissue (SAT) and each sample submitted to sequencing contained tissues from at least 2 mice. The tissue was submitted to collagenase digestion to separate single cells. After collagenase digestion, the cells were strained sequentially from 100, 70, and 40 micron strains to remove any residual tissue or adipocytes. The collected stromal vascular fraction (SVF) was then incubated with a hypotonic buffer to lyse all red blood cells before subjected to scRNA-seq using the 10X Genomics Chromium Single Cell Kit.  Nucleic acid extraction was performed in accordance to UC Davis Genome Center protocols using the 10X Genomics Chromium Single Cell Kit.  Library construction was performed in accordance to UC Davis Genome Center protocols using the 10X Genomics Chromium Single Cell Kit.