Time series analysis of transmesothelial invasion by endometrial stromal and epithelial cells using three-dimensional confocal microscopy.
OBJECTIVE
To evaluate endometrial adhesion and invasion of peritoneal mesothelium.


METHODS
Descriptive study using confocal laser-scanning microscopy.


METHODS
University-based laboratory.


METHODS
Women undergoing surgery for benign conditions.


METHODS
Fluorescence-labeled peritoneal mesothelial cells (PMCs) were grown on coverslips. Fluorescence-labeled endometrial stromal cells (ESCs) and epithelial cells (EECs) and myometrial cells (Myos) were plated on the PMCs. Cultures were examined at 1, 6, 12, and 24-27 hours with differential interference contrast and confocal laser-scanning microscopy.


METHODS
Demonstration of adherence and invasion of endometrial cells through peritoneal mesothelium.


RESULTS
At 1 hour, there was adherence of the ESCs, EECs, and Myos on the perimeter of PMCs. There was no invasion by the Myos. By 6 hours, ESCs and EECs spread over the surface of the PMCs and extended cell processes through PMC junctions. Extension of pseudopodia under the PMCs followed. By 12 hours, there was vacuolization and lifting of PMCs that had been undermined by endometrial cells.


CONCLUSIONS
This is the first time-phase study to demonstrate adherence and the process of invasion of endometrial cells through the mesothelium. The application of three-dimensional confocal laser-scanning microscopy is a novel technique that can be used to further examine mechanisms involved in the pathogenesis of the early endometriotic lesion.
Witz
Craig A
CA
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229-3900, USA. witz@uthscsa.edu
Cho
Sook
S
Centonze
Victoria E
VE
Montoya-Rodriguez
Iris A
IA
Schenken
Robert S
RS
D000328:Adult; D002448:Cell Adhesion / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D004715:Endometriosis / Q000473:pathology*; D004717:Endometrium / Q000166:cytology*; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods; D008857:Microscopy, Interference / Q000379:methods; D010537:Peritoneum / Q000166:cytology; D017154:Stromal Cells / Q000166:cytology; D013997:Time Factors