1. Development and characterization of an endometrial tissue culture model for study of early implantation events 
Tian-Min Ye, M.D., Ph.D., Ronald T.K. Pang, Ph.D., Carmen O.N. Leung, M.Med.Sc., Weimin Liu, Ph.D., William S.B. Yeung, Ph.D. 
Fertility and Sterility 
Volume 98, Issue 6, Pages (December 2012)
DOI: /j.fertnstert
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Culture of endometrial tissue. (A) Diagram showing the setup of the culture system. The photograph shows the placement of the endometrium tissue (arrow) on a metal mesh overlaid with an amniotic membrane matrix (arrowhead). (B) Representative histologic section of day-4 pregnant mouse endometrium cultured on AMM, on MM only, or on LP for 48 hours and negative control (NC) and positive control (PC) of freshly dissected day-4 pregnant mouse endometrium. The apoptotic nuclei in the sections were stained brown. Arrows in PC indicate some of the representative apoptotic signal. LE = luminal epithelium; GE = glandular epithelium; SC = stromal cells. Scale bars = 50 μm. (C) Morphometric parameters and percentage of apoptotic cells in endometrial tissues cultured for 48 and 72 hours. Endometrial tissues from day 6 (Psu-D6) and day 7 (Psu-D7) of pseudopregnancy were used as the corresponding in vivo controls. Box plots show median and 25th–75th percentiles of the corresponding data from five different mice. Error bars indicate the lower and upper extremes, respectively. AD = apoptotic cell density. *P<.05 (ANOVA followed by Dunn's multiple comparison test) vs. the corresponding in vivo controls. N/A = data not available owing to degradation of tissue.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Steroid effect on cultured endometrium. Representative histologic sections of (A) day-4 pregnant endometrium cultured in the medium supplemented with 30% fetal bovine serum plus 0.9 nmol/L E2 plus 63.5 nmol/L P for 72 hours, and (B) freshly dissected endometrial tissue from day-7 pseudopregnant mouse. Scale bars = 50 μm. Insets indicate the higher magnified view of the luminal epithelium with underlying stromal cells (top) and the glandular epithelium (bottom). Scale bars = 20 μm. LE = luminal epithelium; GE = glandular epithelium; SC = stromal cells. (C) Morphometric parameters of endometrial tissues cultured in different steroid environments for 48 and 72 hours. Endometrial tissues from day 6 (Psu-D6) and day 7 (Psu-D7) of pseudopregnancy were used as the corresponding in vivo controls. Box plots show median and 25th–75th percentiles of the corresponding data from five different mice. Error bars indicate the lower and upper extremes, respectively. *P<.05 (ANOVA followed by Dunn's multiple comparison test) vs. the corresponding in vivo controls. N/A = data not available owing to degradation of tissue.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Attachment of blastocyst onto cultured endometrium. Immunohistochemical staining of Oct-4 (A) and CDX-2 (C) on an attached blastocyst in the coculture model, and negative control of Oct-4 (B) and CDX-2 (D). Arrow indicates invasion of the trophoblast through the luminal epithelium. Arrowhead indicates primitive endoderm. LE = luminal epithelium; GE = glandular epithelium; SC = stromal cells. Scale bars = 50 μm. The steroid environment affects the attachment rate on endometrium from day-4 pregnant (E) and day-7 pseudopregnant (F) mice. Embryo attachment was examined after 28 hours of coculture. Those remaining on the endometrium after gentle flushing for three times were considered attached blastocysts. Numbers indicate embryos attached/embryos loaded. *P<.05, χ2 test.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
Decidualization in the cultured endometrium. (A) COX-2, (D) Cx43, and (G) PPARδ expression in day-5 pregnant mouse endometrium in vivo. Positive signal was only detected around the implanting blastocyst. (B) COX-2, (E) Cx43, and (H) PPARδ immunoreactivities were detected in the epithelial or/and stromal cells around the attached embryo in the coculture model, with intensities that decreased with the distance from the attachment site. There was no COX-2 (C), Cx43 (F), or PPARδ (I) expression in the cultured endometrium without blastocyst. (J) Negative control of day-5 pregnant mouse endometrium in vivo. BL = blastocyst; LE = luminal epithelium; GE = glandular epithelium; SC = stromal cells. Scale bars = 200 μm (A, D, and G) and 50 μm (B, C, E, F, H, I, and J).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions