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Hum. Reprod. Advance Access published online on November 17, 2005
Human Reproduction, doi:10.1093/humrep/dei387
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© The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received July 14, 2005
Revised October 4, 2005
Accepted October 7, 2005

Article

Quantification of endometriotic lesions in a murine model by fluorimetric and morphometric analyses

Sylvie Defrère 1, Anne Van Langendonckt 1, Reinaldo González Ramos 1, Mathieu Jouret 1, Marcel Mettlen 2, and Jacques Donnez 1 *

1 Department of Gynaecology, Université Catholique de Louvain, Avenue Hippocrate 10, 1200 Brussels
2 CELL Unit, Université Catholique de Louvain and Christian de Duve Institute of Cellular Pathology, 1200 Brussels, Belgium

* To whom correspondence should be addressed.
Jacques Donnez, E-mail: donnez{at}gyne.ucl.ac.be


  Abstract

BACKGROUND: In animal models of endometriosis, the identification and quantification of lesions originating from human endometrium is often hampered by the small size of the implants and their embedding in murine tissue. The purpose of the present study was to develop two new methods of quantifying endometriosis-like lesions in a nude mouse model: fluorimetry and morphometry. METHODS: Human menstrual endometrium was labelled using a fluorescent tracker, carboxyfluorescein diacetate, succinimidyl ester (CFDA-SE), and transplanted into the pelvic cavity of mice by injection through the peritoneum after performing a cutaneous incision. After 5 days, lesions were recovered by laparotomy. The fluorescence of the recovered endometriotic lesions was measured. Endometrial stroma and glands were immunostained in lesion sections with anti-CD10 and anti-CK22 antibodies, and their surface area was evaluated by morphometric analysis. RESULTS: Fluorescent labelling allows identification of lesions not visible macroscopically. A good correlation was observed between fluorimetry and morphometry (r = 0.88) applied for lesion quantification. CONCLUSIONS: Fluorimetric evaluation combined with morphometric analysis of endometriosis-like lesions allows objective and reliable recording of endometriosis development in a nude mouse model. This quantification method could therefore be useful for future pharmacological and toxicological studies.

Keywords: carboxyfluorescein diacetate, succinimidyl ester/endometriosis/lesion quantification/menstrual endometrium/nude mouse model.
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