fluorimetric and morphometric analyses

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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 flu-orescent 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 recov-ered 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 mac-roscopically. 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 quan-tification method could therefore be useful for future pharmacological and toxicological studies. Key words: carboxyfluorescein diacetate, succinimidyl ester/endometriosis/lesion quantification/menstrual endometrium/nude mouse model

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endometriosis

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