GSM9462790: NC_ATSVF; Mus musculus; RNA-Seq
Adipose tissues were minced into small fragments and enzymatically digested in collagenase type I (1 mg/mL) at 37°C for 30–40 min with gentle agitation. The digested tissue was filtered through a 70-µm cell strainer to remove debris, then centrifuged to separate the stromal vascular fraction (SVF) from mature adipocytes. Red blood cells were lysed using ACK lysis buffer. SVF cells were washed and resuspended in PBS containing 0.04% BSA. Cell viability and concentration were assessed by trypan blue exclusion; samples with viability >85% were used for downstream processing. Single-cell gene-expression libraries were prepared using the Chromium Single Cell 3′ v1 platform (10x Genomics) following the manufacturer's protocol. Approximately 10,000–13,000 cells were loaded per channel to minimize doublets. Library quality was assessed on an Agilent Bioanalyzer. Libraries were sequenced on an Illumina NovaSeq 6000 in paired-end mode (PE150).