Fig. 2: Reconstituting parental tissue-like endometrial features in the EoCs with a trophoblast invasion model. | Nature Communications

Fig. 2: Reconstituting parental tissue-like endometrial features in the EoCs with a trophoblast invasion model.

From: Microengineered patient-derived endometrium-on-a-chip for the evaluation of endometrial receptivity and personalised translational medicine

Fig. 2: Reconstituting parental tissue-like endometrial features in the EoCs with a trophoblast invasion model.

a Schematic of the mouse estrous cycle (proestrus, estrus, metestrus, diestrus), indicating tissue collection points for integration into the EoC. Created in BioRender. Ahn, J. (2025) https://BioRender.com/b09d300b, c Immunofluorescence (IF) images and quantification of ER-β (yellow) with DAPI (blue) in both the EoC (left) and the parental tissues (right) across the estrous cycle (EoC; P vs. D: p = 0.0006). d, e IF and quantification of PR (red) with DAPI (blue) in both the EoC (left) and the parental tissues (right) across the estrous cycle (n = 3 independent mice; one EoC per mouse.) Scale bar: 50 µm. f, g IF of OPN (green) with DAPI (blue) in the EoC and quantification across the estrous cycle (EoC; P vs. E: p < 0.0001, P vs. M: p = 0.0051/ Tissue; P vs. E: p < 0.001, P vs. M: p < 0.0396, E vs. M: p < 0.001, E vs. D: p < 0.001). h, i IF of blood vessel (CD31, red) with DAPI (blue) in the EoC and quantification of blood vessel area (Tissue; P vs. E: p < 0.0013, P vs. M: p < 0.0187, P vs. D: p < 0.0031, E vs. M: p < 0.001, E vs. D: p < 0.001). n = 3 independent mice; one EoC per mouse. Scale bar: 200 µm. j Diagram illustrating trophoblast spheroid aggregation, stromal cell seeding in the EoC, and schematic of the 4-day trophoblast invasion assay. Created in BioRender. Ahn, J. (2025) https://BioRender.com/b09d300k IF images of trophoblast invasion into the stromal layer in the EoC, with F-actin (green), BeWo (red), and DAPI (blue). Scale bar, 50 µm. Quantitative evaluation of the volume of trophoblast spheroid invasion (l), invasion area (m), F-actin volume (n) the number of endometrial stromal cells physically in contact with the trophoblast spheroids (o) over four days (I; D1 vs. D4: p = 0.0031, D2 vs. D4: p = 0.0116/ m; D1 vs. D3: p = 0.0159, D1 vs. D4: p = 0.0008, D2 vs. D3: p = 0.016, D2 vs. D4: p = 0.0008, o; D1 vs. D4: p = 0.0102). n = 3 independent experiments using BeWo spheroids. All data shown as mean ± SD and analysed by one-way ANOVA including P-values (*p < 0.05, **p < 0.01, *** p < 0.001, ****p < 0.0001.) P Proestrus, E Estrus, M Metestrus, D Diestrus. Source data are provided as a Source Data file.

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