Biomedical subjects

Sylvie Defrère

Publications and source records attributed to Sylvie Defrère.

5 recordsLinked to original sources

PubMed2006

Iron overload enhances epithelial cell proliferation in endometriotic lesions induced in a murine model.

BACKGROUND: Iron deposits are characteristic of endometriotic lesions, and pelvic iron concentrations are higher in endometriosis patients than in women without endometriosis. In this study, the effect of iron overload and iron chelation on the development of endometriosis in a murine model was investigated. METHODS: Human menstrual endometrium was injected i.p. into nude mice, either alone (controls) or supplemented with erythrocytes or desferrioxamine (DFO), an iron chelator. After 5 days, the iron load of endometriosis-like lesions and peritoneal macrophages and fluid was evaluated. Lesions were quantified by immunohistochemical morphometry, and their proliferative activity was assessed. RESULTS: Injection of erythrocytes into the pelvic cavity caused iron overload in lesions (P < 0.025) and peritoneal macrophages (P < 0.01) and fluid (P < 0.05), whereas DFO effectively reduced iron status in lesions (P < 0.05) and macrophages (P < 0.01) compared with controls. No difference was observed in the number or surface area of lesions between the three groups. Erythrocytes increased (P < 0.05) and DFO significantly decreased (P < 0.01) the proliferative activity of lesions. CONCLUSIONS: Iron overload does not appear to affect lesion establishment but may contribute to the further growth of endometriosis by promoting cell proliferation of lesions. Iron chelator treatment could therefore be beneficial in endometriosis to prevent iron overload in the pelvic cavity and decrease cellular proliferation of lesions.

Animals↗

PubMed2005

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

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.

Animals↗

PubMed2005

Human endometrial epithelial cells (EEC) constitutively express more intercellular adhesion molecule (ICAM)-1 than endometrial stromal cells (ESC) in culture.

PROBLEM: Intercellular adhesion molecule-1 (ICAM-1) is thought to play an important role in pathophysiological processes in endometrial tissue. The aim of this study was to quantify and compare the expression of ICAM-1 mRNA and protein in cultured endometrial epithelial cells (EEC) versus endometrial stromal cells (ESC). METHOD OF STUDY: EEC and ESC were isolated from human endometrium and cultured. At confluency, ICAM-1 mRNA was measured by real-time reverse transcriptase-polymerase chain reaction, the membrane-bound form (mICAM-1) by immunocytodensitometry, and the soluble form (sICAM-1) by enzyme-linked immunosorbent assay. RESULTS: At the transcriptional level, we observed a 1.7-fold increase in ICAM-1 expression in EEC versus ESC. mICAM-1 immunostaining in EEC [cell-relative membrane-bound signal-specific optical density (CR-M-SOD): 0.056 +/- 0.05] was stronger (P < 0.05) than in ESC (CR-M-SOD: 0.009 +/- 0.07). EEC were found to secrete more (P < 0.01) sICAM-1 ([sICAM-1] = 15.59 +/- 2.96 ng/mL) than ESC ([sICAM-1] = 5.14 +/- 2.61 ng/mL). CONCLUSIONS: Cultured EEC constitutively express significantly more ICAM-1 mRNA and protein than ESC.

Adult↗

PubMed2004

Ficoll density gradient method for recovery of isolated human ovarian primordial follicles.

OBJECTIVE: To develop a simple and efficient technique to allow rapid recovery of a maximum number of good quality isolated follicles. DESIGN: Prospective experimental study. SETTING: Academic research unit of the department of gynecology in a university hospital. PATIENT(S): Biopsies were obtained from five women (between 26 and 31 years of age). INTERVENTION(S): Biopsies were cut with a tissue sectioner. Enzymatic digestion was performed in a collagenase solution for 90 min at 37 degrees C. The follicles were recovered using a discontinuous Ficoll density gradient method. MAIN OUTCOME MEASURE(S): The number of follicles present in the interface layers of Ficoll gradient was quantified. Follicular viability of these recovered follicles was assessed with live-dead stains, using calcein-AM and ethidium homodimer-I. RESULT(S): Out of a total of 6,811 recovered follicles, we found 63% (n = 4,201) at the medium-1.06 Ficoll interface and 36.9% (n = 2,590) at the 1.06-1.09 Ficoll interface, which represents 99.9% of total recovered follicles. Analysis by vital fluorescent staining showed that 95.8% of the follicles treated with Ficoll were totally viable. CONCLUSION(S): The Ficoll density gradient method allows us to maximize the recovery of isolated human ovarian follicles and minimize the manipulation time while maintaining high follicular viability.

Adult↗

PubMed2004

Freeze-thawing intact human ovary with its vascular pedicle with a passive cooling device.

OBJECTIVE: To test the feasibility of freezing intact human ovary using a passive cooling device. DESIGN: Prospective experimental study. SETTING: Academic research unit of the department of gynecology in a university hospital. PATIENT(S): Ovaries were obtained from three women who were between 29 and 36 years of age. INTERVENTION(S): Ovarian perfusion with cryoprotective solution and slow freezing with a cryofreezing container (5100 Cryo 1 degrees C Freezing Container; Nalgene, VWR, Belgium). Rapid thawing of ovaries by perfusion and bathing with decreased sucrose gradient. MAIN OUTCOME MEASURE(S): Viability of follicles, stromal cells, and vascular components was assessed with live-dead stains in freshly removed ovary, after cryoprotectant exposure before freezing, and after thawing. Histological morphology was assessed at these three different times. RESULT(S): The percentage of live follicles was 99.4% in fresh tissue, 98.1% after cryoprotectant exposure, and 75.1% after thawing. Viability assessment showed live stromal cells and small vessels after thawing. On histological evaluation, the morphology of follicles and cortical and medullar tissue was similar in all three groups. CONCLUSION(S): We described the cryopreservation, using an accessible protocol, of intact human ovary with its vascular pedicle and proved high survival rates of follicles, small vessels, and stromal cells and a normal histological structure in all the ovarian components after thawing using our protocol.

Adult↗