GSM9693284: SMSCs from Zhedong White goose leg muscle , embryonic days 13 (E23), scRNA-seq; Anser cygnoides; RNA-Seq
Satellite cells were isolated and purified according to our established protocol [9]. Briefly, muscle tissues were minced and digested in high-glucose DMEM with 2 mg/mL Dispase II and 4 mg/mL Collagenase II (Roche, Gibco) at 37°C for 50 minutes. Digestion was stopped using DMEM with 10% FBS. The cell suspension was filtered through a 70 μm strainer, centrifuged at 350× g for 8 minutes, and red blood cells lysed with ACK buffer. Differential plating was performed twice to enrich SMSCs, exploiting fibroblast attachment for one hour each time. Purity was confirmed by Pax7 immunofluorescence staining [9]. Enriched SMSCs were cryopreserved in liquid nitrogen for subsequent scRNA-seq. Thawed SMSCs were filtered through a 40 μm strainer, centrifuged at 300× g for 5 minutes at 4°C, and resuspended in PBS with 0.04% BSA. Only samples with >85% via-bility (trypan blue exclusion) and 400–1,200 cells/μL were processed for single-cell cap-ture. Chromium Single Cell 3′ Library Kit (10× Genomics) was used for cell capture, barcoding, and cDNA synthesis following manufacturer's protocols . Libraries were quantified using a Bioanalyzer 2100 and Qubit, then sequenced on Illumina NovaSeq platforms (PE 150 bp). Library preparation and sequencing were conducted at Shanghai Personal Biotechnology Co., Ltd. and Shanghai Majorbio Bio-pharm Technology Co., Ltd..
GEO
GSM9693284_r1