为了研究经黄体酮处理的不孕和生育子宫内膜类器官之间的转录组谱是否存在差异。从 14 名不孕女性和 7 名生育女性中获取子宫内膜活检组织，然后从分离的上皮细胞中生成类器官。为了模拟分泌期，类器官依次用 17β-雌二醇 (E2) 和孕酮 (P4) 处理，并进行 RNA 测序。使用DESeq2（lfcThreshold = 0，log2 Fold Change ≥ 1.0或 ≤ -1.0）鉴定差异表达基因（DEG），并生成主成分分析（PCA）图。通过过度代表性分析和基因集富集分析（GSEA）进行功能富集分析。为了在功能上评估增殖，在类器官分化之前（T0）和之后（T1）进行 OrganoSeg 表面测量，并计算 T1/T0 比率以确定增殖率。尽管 PCA 图没有显示可育和不育的清晰聚类与可育的类器官相比，在不育样本中检测到 363 个显着的 DEG（129 个上调和 234 个下调）。主要是细胞周期过程在不育类器官中高度富集。因此，我们假设与可育类器官相比，不育类器官分化过程中的增殖活性可能更高。然而，这无法通过细胞表面测量来验证。这项研究表明，与可育的类器官相比，E2/P4 处理的不育子宫内膜类器官的细胞周期过程更加丰富。这可能反映了与可育类器官相比，分化的不育类器官中子宫内膜上皮细胞的增殖活性持续较高。为了证实这一假设，需要进一步研究。© 2024。作者。

To investigate whether the transcriptome profile differs between progesterone-treated infertile and fertile endometrial organoids.Endometrial biopsies were obtained from 14 infertile and seven fertile women, after which organoids were generated from isolated epithelial cells. To mimic the secretory phase, organoids were sequentially treated with 17β-estradiol (E2) and progesterone (P4) and subjected to RNA sequencing. Differentially expressed genes (DEGs) were identified using DESeq2 (lfcThreshold = 0, log2 Fold Change ≥ 1.0 or ≤ -1.0), and a principal component analysis (PCA) plot was generated. Functional enrichment analysis was performed by overrepresentation analysis and Gene Set Enrichment Analysis (GSEA). To functionally assess proliferation, OrganoSeg surface measurements were performed before (T0) and after (T1) differentiation of organoids, and T1/T0 ratios were calculated to determine the proliferation rate.Although the PCA plot did not show clear clustering of the fertile and infertile samples, 363 significant DEGs (129 upregulated and 234 downregulated) were detected in infertile compared to fertile organoids. Mainly cell cycle processes were highly enriched in infertile organoids. Thus, we hypothesised that proliferative activity during differentiation may be higher in infertile organoids compared to fertile organoids. However, this could not be validated by cell surface measurements.This study revealed that cell cycle processes were enriched in E2/P4-treated infertile endometrial organoids as compared to fertile organoids. This could reflect persistently higher proliferative activity of the endometrial epithelial cells in differentiated infertile organoids compared to fertile organoids. To confirm this hypothesis, further studies are warranted.© 2024. The Author(s).