title: GSM9162114: Decidua, C02, first trimester, replicate 2; Homo sapiens; RNA-Seq
library_construction_protocol: Human decidual tissues from first-trimester elective surgical terminations were collected and immediately processed. Tissues were enzymatically dissociated using the Human Tumor Dissociation Kit (Miltenyi Biotec, 130-095-929) and the gentleMACS™ Octo Dissociator (Miltenyi Biotec, 130-096-427) following the manufacturer's instructions. The resulting cell suspension was filtered through a 70 μm cell strainer (Miltenyi Biotec, 130-101-262), followed by red blood cell lysis using RBC Lysis Buffer (Miltenyi Biotec, 130-094-183). Dead cells were removed and cell viability was assessed using trypan blue exclusion. Samples with cell viability above 85% were used for single-cell library preparation. Single-cell RNA-seq libraries were constructed using the 10x Genomics Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (10x Genomics, 1000121) according to the manufacturer's protocol. Briefly, viable single-cell suspensions were loaded onto the Chromium Controller to generate Gel Bead-in-Emulsions (GEMs), enabling barcoding of individual cells. Reverse transcription was performed within droplets, followed by cDNA amplification and library preparation steps including fragmentation, end repair, A-tailing, adaptor ligation, and sample indexing. The final libraries were quantified, pooled, and subjected to paired-end sequencing.