1. Secretion of monocyte chemotactic protein-1 by human uterine epithelium directs monocyte migration in culture 
Richard A. Meter, M.D., Charles R. Wira, Ph.D., John V. Fahey, Ph.D. 
Fertility and Sterility 
Volume 84, Issue 1, Pages (July 2005)
DOI: /j.fertnstert
Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

2. FIGURE 1
Chemotaxis assay of THP-1 monocytes to rMCP-1 in apical, basolateral, and both chambers of transwells. 2 ×105 THP-1 monocytes were placed in all apical compartments of 5.0-μm pore transwells. Monocyte chemotactic protein-1 at 25 ng/mL was placed in the basolateral chambers only (positive control) in one group of transwells, the apical chambers only in a second group, and in both chambers in a third group. Monocytes that migrated through the pores were counted after a 2-hour incubation. The mean of triplicate transwells ± SEM is shown. ***Significant difference from negative control (no MCP-1), P<.001.
Meter. Uterine epithelium and chemotaxis. Fertil Steril 2005.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

3. FIGURE 2
Dose response of THP-1 monocytes to rMCP-1 in chemotaxis assay. 1×105 THP-1 monocytes were placed in all apical chambers of 5.0-μm pore transwells above varying concentrations of rMCP-1 in the basolateral chambers. Monocytes that migrated through the pores were counted after a 2-hour incubation. The mean of triplicate transwells ± SEM is shown. **Significant difference from negative control, P<.01; ***significant difference from negative control, P<.001.
Meter. Uterine epithelium and chemotaxis. Fertil Steril 2005.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

4. FIGURE 3
Chemotaxis assay of THP-1 monocytes to apical and basolateral culture media from endometrial epithelial cells grown in culture. Endometrial epithelial cells were first grown to high TER on cell culture inserts. After 48 hours in fresh culture media, the cell culture media was collected from the apical and basolateral compartments of the inserts. These epithelial exposed media were then placed in the bottom chambers of 5.0-μm pore transwells. 1 × 105 THP-1 monocytes were placed in all top chambers. Monocyte chemotactic protein-1 at 25 ng/mL was placed in the bottom chamber of the positive controls. THP-1 monocytes that migrated through the pores were counted after a 2-hour incubation. The mean of triplicate transwells ± SEM is shown. *Significant difference from negative control, P<.05; **significant difference from negative control, P<.01.
Meter. Uterine epithelium and chemotaxis. Fertil Steril 2005.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

5. FIGURE 4
Luminex-Bioplex analysis of MCP-1 from endometrial epithelial cell culture of six hysterectomy surgical specimens. Apical and basolateral cell culture media were obtained after growing the cell cultures to high TER. The samples were then analyzed by Luminex for MCP-1. The mean of quadruplicate cultures ± SEM is shown.
Meter. Uterine epithelium and chemotaxis. Fertil Steril 2005.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

6. FIGURE 5
Chemotaxis of THP-1 monocytes to endometrial epithelial culture media is substantially eliminated after incubation of the epithelial culture media with anti-hMCP-1 antibody. Surgically derived endometrial epithelial cells from patient 2694 were cultured and grown to high TER on cell culture inserts. Forty-eight hour apical culture media was obtained and incubated for 1.5 hours with polyclonal anti-hMCP-1 antibody, then placed in the bottom chambers of 5.0-μm pore transwells. The same procedure was performed using the respective isotype antibody. 1×105 THP-1 monocytes were placed in all top chambers. Monocyte chemotactic protein-1 at 25 ng/mL was placed in the bottom chamber of the positive controls. Monocytes that migrated through the pores were counted after a 2-hour incubation. The mean of triplicate transwells ± SEM is shown. **Significant difference compared with positive control, P<.01; ***significant difference compared with positive control, P<.001.
Meter. Uterine epithelium and chemotaxis. Fertil Steril 2005.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

7. FIGURE 6
Chemotaxis of THP-1 monocytes to endometrial epithelial culture media from additional patients after incubation with anti-hMCP-1 antibody. Apical and basolateral culture media were obtained from surgical patient-derived endometrial epithelial cell cultures from 3 additional surgical patients. The apical and basolateral culture media were combined for each patient to produce the volumes required for the assay. After 1.5-hour incubation with monoclonal anti-hMCP-1 antibody, the culture media were placed in the bottom chambers of 5.0-μm pore transwells. The same was done using the respective isotype antibody. 1×105 THP-1 monocytes were placed in all top chambers. Monocyte chemotactic protein-1 at 10 ng/mL was placed in the bottom chamber of the positive controls. Monocytes that migrated through the pores were counted after a 2-hour incubation. The mean of triplicate transwells ± SEM is shown. *Significant difference compared with positive control, P<.05; ***significant difference compared with positive control, P<.001.
Meter. Uterine epithelium and chemotaxis. Fertil Steril 2005.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions