绒毛膜癌球体与接受型和非接受型子宫内膜上皮细胞系相互作用的转录组分析：一种用于人类着床的体外模型。
Transcriptomic analysis of the interaction of choriocarcinoma spheroids with receptive vs. non-receptive endometrial epithelium cell lines: an in vitro model for human implantation.
机构信息
Clínica EUGIN, Travessera de les Corts 322, 08029, Barcelona, Spain.
Facultat de Biociències, Departament de Biologia Cel·lular, de Fisiologia i d'Immunologia, Universitat Autònoma de Barcelona, Barcelona, Spain.
出版信息
J Assist Reprod Genet. 2019 May;36(5):857-873. doi: 10.1007/s10815-019-01442-9. Epub 2019 Apr 10.
PURPOSE
Several in vitro systems have been reported to model human implantation; however, the molecular dynamics of the trophoblast vs. the epithelial substrate during attachment have not been described. We have established an in vitro model which allowed us to dissect the transcriptional responses of the trophoblast and the receptive vs. non-receptive epithelium after co-culture.
METHODS
We established an in vitro system based on co-culture of (a) immortalized cells representing receptive (Ishikawa) or non-receptive (HEC-1-A) endometrial epithelium with (b) spheroids of a trophoblastic cell line (JEG-3) modified to express green fluorescent protein (GFP). After 48 h of co-culture, GFP+ (trophoblast cells) and GFP- cell fractions (receptive or non-receptive epithelial cells) were isolated by fluorescence-activated flow cytometry (FACS) and subjected to RNA-seq profiling and gene set enrichment analysis (GSEA).
RESULTS
Compared to HEC-1-A, the trophoblast challenge to Ishikawa cells differentially regulated the expression of 495 genes, which mainly involved cell adhesion and extracellular matrix (ECM) molecules. GSEA revealed enrichment of pathways related to cell division, cell cycle regulation, and metabolism in the Ishikawa substrate. Comparing the gene expression profile of trophoblast spheroids revealed that 1877 and 323 genes were upregulated or downregulated when co-cultured on Ishikawa substrates (compared to HEC-1-A), respectively. Pathways favorable to development, including tissue remodeling, organogenesis, and angiogenesis, were enhanced in the trophoblast compartment after co-culture of spheroids with receptive epithelium. By contrast, the co-culture with less receptive epithelium enriched pathways mainly related to trophoblast cell proliferation and cell cycle regulation.
CONCLUSIONS
Endometrial receptivity requires a transcriptional signature that determines the trophoblast response and drives attachment.
目的
已经有几种体外系统被报道可以模拟人类着床；然而，滋养层与附着时的上皮基质之间的分子动力学尚未描述。我们建立了一种体外模型，使我们能够在共培养后分析滋养层和接受性与非接受性上皮的转录反应。
方法
我们建立了一种基于共培养的体外系统，其中包括(a) 代表接受性（Ishikawa）或非接受性（HEC-1-A）子宫内膜上皮的永生化细胞，以及(b) 表达绿色荧光蛋白（GFP）的滋养层细胞系（JEG-3）的球体。共培养 48 小时后，通过荧光激活流式细胞术（FACS）分离 GFP+（滋养层细胞）和 GFP-细胞群（接受性或非接受性上皮细胞），并进行 RNA-seq 分析和基因集富集分析（GSEA）。
结果
与 HEC-1-A 相比，滋养层对 Ishikawa 细胞的挑战差异调节了 495 个基因的表达，这些基因主要涉及细胞黏附和细胞外基质（ECM）分子。GSEA 显示，在 Ishikawa 基质中，与细胞分裂、细胞周期调控和代谢相关的途径富集。比较滋养层球体的基因表达谱显示，当与 Ishikawa 基质共培养时，分别有 1877 和 323 个基因上调或下调（与 HEC-1-A 相比）。有利于发育的途径，包括组织重塑、器官发生和血管生成，在与接受性上皮共培养的滋养层中得到增强。相比之下，与接受性较低的上皮共培养富集了主要与滋养层细胞增殖和细胞周期调控相关的途径。
结论
子宫内膜容受性需要一个转录特征，该特征决定了滋养层的反应并驱动附着。
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