Homo sapiens Raw sequence reads

All human endometrial samples were obtained from patients at the obstetrics and gynecology hospital of Fudan University in China. Normal endometrium was obtained from patients who are subjected to hysterectomy due to uterine fibroids or early cervical cancers. Samples of precancerous endometrial lesions and EEC were obtained from patients who received hysterectomy because of the aforementioned diseases. In order to prevent the effect of age on endometrium, endometrial samples were only obtained from women of 44 to 55 years old. The collected endometrial tissues were mechanically cut by scalpels as small as possible, followed by dissociating in 20 ml of endometrial digestion medium with gentle shaking at 37 C for 1-2 h. Pipetted up and down every 20 min. Till most cells in single cell status, suspended the digestion and filtered the digested cells with 40um cell sieves and obtained single cell suspension. The single cell suspension was centrifuged at 800rpm for 3min and then lysed red blood cell. For scRNA-seq, the percentage of the live single cells (counted by Typan blue) was over 80% in total endometrial cells before manufacturing in the single-cell-A-Chip (10X Genomics). Then, the chip with endometrial cells was loaded in a 10X Chromium single-cell instrument (10X Genomics). The following barcoding, cDNA synthesis and library construction were followed by standard manufacturer`s instructions. The qualified libraries were applied to the Illumina NovaSeq6000 platform for PE150 sequencing.本研究所有人类子宫内膜样本均取自中国复旦大学附属妇产科医院的患者。正常子宫内膜样本取自因子宫肌瘤或早期宫颈癌接受子宫切除术的患者；子宫内膜癌前病变及子宫内膜样腺癌（Endometrial Endometrioid Carcinoma, EEC）样本取自因上述疾病接受子宫切除术的患者。为排除年龄对子宫内膜状态的影响，本研究仅纳入44~55岁女性的子宫内膜样本。收集的子宫内膜组织先用手术刀尽可能精细地机械切割，随后置于20 mL子宫内膜消化培养基中，于37℃轻柔振摇消化1~2小时，每20分钟上下吹打一次。待大部分细胞呈单细胞状态后，终止消化反应，将消化后的细胞通过40 μm细胞筛过滤，获得单细胞悬液。将所得单细胞悬液以800 rpm离心3分钟，随后进行红细胞裂解处理。对于单细胞RNA测序（single-cell RNA sequencing, scRNA-seq），在使用单细胞A芯片（single-cell-A-Chip，10X Genomics）进行制备前，经台盼蓝（Trypan Blue）染色计数显示，活单细胞占总子宫内膜细胞的比例需超过80%。随后将装载有子宫内膜细胞的芯片置入10X Chromium单细胞仪器（10X Genomics）中。后续的条形码标记、cDNA合成及文库构建均严格遵循厂商标准操作流程。合格的文库将被上机至Illumina NovaSeq6000平台，进行PE150双端测序。