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Time Series Analysis of Transmesothelial Invasion by Endometrial Stromal and Epithelial Cells Using Three-dimensional Confocal Microscopy

Published online by Cambridge University Press:  02 July 2020

CA Witz
Affiliation:
Departments of Obstetrics and Gynecology, The University of Texas Health Science Center at San Antonio, San Antonio, TX
S Cho
Affiliation:
Departments of Obstetrics and Gynecology, The University of Texas Health Science Center at San Antonio, San Antonio, TX
VE Centonze
Affiliation:
Cellular and Structural Biology, The University of Texas Health Science Center at San Antonio, San Antonio, TX
IA Montoya-Rodriguez
Affiliation:
Departments of Obstetrics and Gynecology, The University of Texas Health Science Center at San Antonio, San Antonio, TX
RS Schenken
Affiliation:
Departments of Obstetrics and Gynecology, The University of Texas Health Science Center at San Antonio, San Antonio, TX
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Abstract

Using human peritoneal explants, we have previously demonstrated that endometrial stromal cells (ESCs) and endometrial epithelial cells (EECs) attach to intact mesothelium. Attachment occurs within one hour and mesothelial invasion occurs within 18 hours (Figure 1). We have also demonstrated that, in vivo, the mesothelium overlies a continuous layer of collagen IV (Col IV).

More recently we have used CLSM, to study the mechanism and time course of ESC and EEC attachment and invasion through mesothelial monolayers. in these studies, CellTracker® dyes were used to label cells. Mesothelial cells were labeled with chloromethylbenzoylaminotetramethylrhodamine (CellTracker Orange). Mesothelial cells were then plated on human collagen IV coated, laser etched coverslips. Mesothelial cells were cultured to subconfluence. ESCs and EECs, labeled with chloromethylfluorscein diacetate (CellTracker Green) were plated on the mesothelial monolayers. Cultures were examined at 1, 6, 12 and 24 hours with simultaneous differential interference contrast and CLSM.

Type
The Cell Biology of Cancer (Organized by J. Jerome and B. Gunning)
Copyright
Copyright © Microscopy Society of America 2001

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References

1.Witz, CA et al. Fertil Steril 2001;74:385-90.CrossRefGoogle Scholar
2.Witz, CA et al. Fertil Steril 1999;71:5660.CrossRefGoogle Scholar