GSM7623269: MI+PUR-AEC ,sc-RNA seq; Rattus norvegicus; RNA-Seq
Tissue collection and CD45+ cells isolation Left ventricles of hearts from the MI and PUR-hAESC groups were collected and minced into fine pieces (including the infarcted and peri-infarcted regions). Tissues were enzymatically digested with collagenase A and elastase at 37°C rotationally. After dissociation, cell suspensions were passed through a 30 µm cell strainer, followed by red blood cell lysis. For sorting of all the immune cells (CD45+), the digested suspensions were stained with Propidium Iodide  and FITC anti- rat CD45 antibody , and sorted on a FACS Aria III (BD Biosciences, Franklin Lakes, NJ) with a 100 µm nozzle. CD45+ cells were sorted using FACS from single-cell suspensions of hearts of MI- or PUR-hAESC-operated rats 1 week after surgery. After washing and resuspending in 0.5mL cold PBS, purified CD45+ cells were processed in the Chromium single cell gene expression platform (10x Genomics). Briefly, Single Cell 3′ Reagent Kit v2 was used to generate droplets encapsulating single cell and barcoded beads. Following capture and lysis, reverse transcription was performed in a thermal cycler (ProFlex PCR). cDNA was synthesized and amplified for 14 cycles. The amplified-cDNA was fragmented, ligated with adapter and sample index, and selected with SPRI beads (Beckman). Sequencing was performed on the NovaSeq 6000 Illumina sequencing platform. Bioinformatic analysis was performed using the OmicStudio tools at https://www.omicstudio.cn/tool. scANS-seq