title: GSM6970370: Patient 1, scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: After the biopsy, the endometrial tissues were processed immediately. The two-stage dissociation protocol was performed. Briefly, the tissues were washed with DMEM/F12 medium (Gibco) and subsequently minced into small pieces. Then, the minced tissues were dissociated in DMEM/F12 supplemented with 1% Penicillin-Streptomycin (Gibco, 15140122), 0.4 mg/mL Collagenase V (Sigma, C9263), 1.25 U/mL Dispase II (Sigma, D4693) at 37 °C for 1 h with agitation. The supernatant was collected as the stromal fibroblast-enriched suspension. The pellet was the glandular and luminal fragments, which was further dissociated in TrypLE Select (Gibco, 12563-011) at 37 °C for 20 min. The cells were collected as the epithelium-enriched portion. Cell counting and quality control were performed following the manufacturer's instructions. Then, the two portions were mixed and loaded into Chromium microfluidic chips (Chromium Next GEM Chip G Single Cell Kit, 10X Genomics) with 3' chemistry and barcoded with a 10×Chromium Controller. RNA from the barcoded cells was subsequently reverse-transcribed to generate barcoded cDNA. Next, sequencing libraries were constructed with reagents from a Chromium Single Cell 3' v3.1 reagent kit (10X Genomics) and Dynabeads™ MyOne™ SILANE (Life, 37005D) according to the manufacturer's instructions. The single-cell libraries were sequenced in paired-end reads on a NovaSeq 6000 platform (Illumina).