GSM9676228: cardiac cells, sham control, collected at day4; Gallus gallus; RNA-Seq
Cardiac neural crest cell (CNCC)-derived cells were isolated from chick embryos collected at days 4 to 7 from normal control and left atrial ligation (LAL) conditions. Embryonic cardiac tissues containing CNCC derivatives were dissected and dissociated into single-cell suspensions using enzymatic digestion and gentle mechanical trituration. Cell suspensions were filtered to remove clumps and debris, and viable fluorescently labeled cells were enriched by fluorescence-activated cell sorting based on H2B-RFP signal. Sorted cells were collected in buffer compatible with single-cell capture and immediately processed for 10x Genomics library preparation. Single-cell libraries were prepared using the 10x Genomics Chromium Single Cell 3′ Gene Expression workflow following the manufacturer's protocol. Briefly, sorted single cells were partitioned into nanoliter-scale gel bead-in-emulsions (GEMs), where cell lysis and reverse transcription were performed. cDNA molecules were labeled with cell barcodes and unique molecular identifiers (UMIs), followed by cDNA amplification, purification, and construction of sequencing-ready libraries. Libraries were sequenced and the resulting reads were processed using Cell Ranger to generate raw and filtered feature-barcode matrices. Downstream analysis was performed in R using Seurat.