1. A tissue-engineered human endometrial stroma that responds to cues for secretory differentiation, decidualization, and menstruation 
Stacey C. Schutte, Ph.D., Robert N. Taylor, M.D., Ph.D. 
Fertility and Sterility 
Volume 97, Issue 4, Pages (April 2012)
DOI: /j.fertnstert
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Change in human endometrial stromal cell morphology in response to hormone treatment. (A) Images of the cells in two-dimensional (2D) with phase contrast or three-dimensional (3D) histologic sections stained with hematoxylin and eosin. Scale bars represent 50 μm. Only images of the early secretory (ES) and decidual (D) conditions are shown, as there are no visible differences noticeable between early secretory, late secretory (LS), and menstruation/steroid withdrawal (M) conditions (n = 6). (B) Cellular (2D) and nuclear (3D) shape indexes calculated from cells and nuclei in six samples to quantify morphologic differences. ∗P<.05 compared with all other conditions within the culture condition (2D or 3D). In 2D culture, decidualization induced a rounder cell shape as compared with the other hormone treatments. In 3D culture, the early secretory phase nuclei were statistically significantly flatter than those in the other hormone treatments, but the late secretory, decidual, and menstruation conditions were not statistically significantly different.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Enzyme-linked immunosorbent assay (ELISA) performed to determine the secretion of decidualization biomarkers (A) prolactin (PRL), (B) insulin-like growth factor binding protein 1 (IGFBP-1), and (C) vascular endothelial growth factor (VEGF), which has been shown to increase during stromal differentiation (n = 6). The results were consistent across all three proteins: in two-dimensional (2D) and three-dimensional (3D) cultures, cells secreted statistically significantly higher amounts of these biomarkers when given the decidualization (D) hormones. The late secretory (LS) stimulus responses were not statistically significantly different from any of the other conditions in 3D culture.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Visible degradation of the three-dimensional (3D) tissues after steroid withdrawal. (A) Images of the two-dimensional (2D) and 3D cultures at day 10. Scale bars indicate 50 μm in the 2D frames and 20 μm in the 3D frames. Examples of late secretory (LS) and menstrual/steroid withdrawal (M) conditions are shown, where the menstrual condition is the only one that showed evidence of breakdown in 3D tissues. (B) Increase in active collagenase seen after steroid withdrawal in 3D tissues, but no statistically significantly affect in 2D cultures (n = 6). Gelatin zymography was performed to visualize latent and active gelatinases (n = 5). (C) Matrix metalloproteinase (MMP) 2 and MMP-9 identified by their molecular weights. The lanes from left to right are 3D: early secretory (ES), late secretory (LS), decidualization (D), and menstrual (M); followed by 2D: ES, LS, D, M. The menstrual conditions led to increased MMP-2 activation in 3D culture, but no differences in 2D culture.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
Subtle differences after hormone treatments in stromal cell numbers as indicated by DNA content. The content for the three-dimensional (3D) was normalized to dry weight to take into consideration differences in hydrogel size. The only significant finding was the lower DNA content detected in the two-dimensional (2D) cultures exposed to decidualization hormones (n = 6).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions