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. 2021 Feb 25;9:616893. doi: 10.3389/fcell.2021.616893

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Copyright © 2021 Eremichev, Kulebyakina, Alexandrushkina, Nimiritsky, Basalova, Grigorieva, Egiazaryan, Dyikanov, Tkachuk and Makarevich.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Tissue-specific features of extracellular matrix (ECM) spatial organization in stromal cells treated by menstrual discharge serum (MDS). (A) Treatment with MDS induces formation of “hills” in dermis stromal cells (DermSCs) and adipose tissue stromal cells (AdipoSCs), but not in endometrium stromal cell (EndoSC) cultures. Vinculin was used as a background labeling antigen to visualize stromal cells; nuclei stained by 4′,6-diamidino-2-phenylindole (DAPI). Confocal microscopy with side projection of resliced z-stacks. Scale bar 50 μm. (B) After MDS treatment, spatial organization of ECM is characterized by superficial localization of ED-A fibronectin above collagen I in DermSCs and AdipoSCs, while EndoSCs lack this feature, displaying homogeneous deposition of these ECM components. Immunofluorescent staining without permeabilization. Side projection of resliced z-stacks. Scale bar 10 μm; confocal microscopy. Whisker-box plots present the distribution of collagen I and ED-A fibronectin localization. Wilcoxon signed rank test was used for statistical analysis; ^∗p < 0.05. (C) Higher level of collagen I and ED-A fibronectin colocalization (Pearson’s correlation coefficient, R) in EndoSC than in DermSC and AdipoSC in MDS-treated cultures. 3D reconstruction of collagen I and ED-A fibronectin deposition pattern. Confocal microscopy.