GSM8723601: ADRCs, Female no. 03, scRNAseq; Homo sapiens; RNA-Seq
ADRCs were isolated from human lipoaspirates using an automated processing Celution® 800/IV device (Lorem Cytori Therapeutics).To enable single-cell RNA sequencing (scRNA-seq), at least 1e+6 ADRCs from each of 14 subjects (7 males and 7 females) were methanol-fixed and stored at -80°C. Briefly, the cells were resuspended in 500 µl PBS with 1% BSA and 1 U/µl RNAsin PLUS RNase Inhibitor (Promega) and incubated on ice for 5 min to ensure the integrity of the RNA in the cells. The cells were passed through a 40 µm Flowmi® Cell strainer (VWR) to create a single-cell suspension. The cells were centrifuged at 400g for 3 min at 4°C, and washed in 1 ml RNase-free PBS and subsequently centrifuged at 400g for 3 min at 4°C, before the cell pellet was resuspended in 200 µl RNase-free PBS and 800 µl ice-cold methanol gently added dropwise to the cells. The cells were fixed for 30 min at -20°C and subsequently stored at -80°C until use. Following rehydration, 8,000 cells were loaded onto the 10x Genomics Chromium controller (10X Genomics). Libraries were prepared according to the instructions of the manufacturer using the 10x Genomics Single-Cell 3' v2 or v3, Chromium Single Cell B Chip Kit, 48 runs (10X Genomics) and sequenced on an Illumina NovaSeq 6000 System (10X Genomics).