To study adhesion between endometrial tissue and peritoneum using intact amniotic membranes as an in vitro model.
A cell biologic and immunohistochemical study.
Tertiary-care university medical ...center.
Ten patients with regular and ovulatory cycles.
Mechanically dissected tissue fragments from endometrial biopsies were cultured on either side of intact amniotic membranes. Also, the carcinoma cell lines RL95–2 and AN3CA were used.
The adhesion of endometrial fragments and of endometrial carcinoma cell lines to amniotic membrane was studied in vitro and evaluated using frozen sections. The composition of the extracellular matrix and the presence of intermediate filament proteins of amniotic membrane were determined using immunohistochemistry and compared with those of normal peritoneum.
Peritoneum and amniotic membrane were similar with respect to expression of cytokeratins in epithelial lining and of extracellular matrix (ECM) components. The endometrial fragments did not adhere to the intact epithelial side of the amniotic membrane. In contrast, adhesion did occur to the nonepithelial side of the amnion. The carcinoma cell lines RL95–2 and AN3CA adhered to either side of intact amniotic membranes.
An intact epithelial lining prevents adhesion of endometrial fragments to the ECM of amniotic membranes in vitro. An intact epithelium could be an important defense mechanism in preventing initial adhesion of retrogradely shed endometrium fragments to peritoneum.
The chick embryo chorioallantoic membrane (CAM) is an established in vivo angiogenesis assay. The aim of our study was to assess the angiogenic properties of endometrium and to quantitate the ...vascular response in an accurate way. Samples of proliferative endometrium (n = 17) and control mouse skin tissue (n = 8) were explanted onto the CAM at day 10 of incubation. Additional controls consisted of normal unmanipulated CAM (n = 12). Four days after grafting, photographs of the explant and the surrounding area were taken in ovo to measure the vascular density index (VDI). The VDI is a stereological estimate of vessel number and length, which was obtained by counting the intersections of vessels with a circular grid superimposed on a computerized image. Endometrium caused a significant increase in VDI as compared to both unmanipulated CAM (p < 0.001) and skin tissue as a control (p < 0.007). The intra-observer variability was 5.2%. This study demonstrates that the CAM assay is a suitable model to assess the angiogenic properties of endometrium. Furthermore, it allows detailed quantitation of the vascular response in an objective and reproducible way. Our findings suggest the CAM to be a promising model to study the role of angiogenesis in both normal human endometrium and diseases involving the endometrium.
Research Priorities for Endometriosis Rogers, Peter A W; Adamson, G David; Al-Jefout, Moamar ...
Reproductive sciences (Thousand Oaks, Calif.),
02/2017, Letnik:
24, Številka:
2
Journal Article
Recenzirano
The 3rd International Consensus Workshop on Research Priorities in Endometriosis was held in São Paulo on May 4, 2014, following the 12th World Congress on Endometriosis. The workshop was attended by ...60 participants from 19 countries and was divided into 5 main sessions covering pathogenesis/pathophysiology, symptoms, diagnosis/classification/prognosis, disease/symptom management, and research policy. This research priorities consensus statement builds on earlier efforts to develop research directions for endometriosis. Of the 56 research recommendations from the 2011 meeting in Montpellier, a total of 41 remained unchanged, 13 were updated, and 2 were deemed to be completed. Fifty-three new research recommendations were made at the 2014 meeting in Sao Paulo, which in addition to the 13 updated recommendations resulted in a total of 66 new recommendations for research. The research recommendations published herein, as well as those from the 2 previous papers from international consensus workshops, are an attempt to promote high-quality research in endometriosis by identifying and agreeing on key issues that require investigation. New areas included in the 2014 recommendations include infertility, patient stratification, and research in emerging nations, in addition to an increased focus on translational research. A revised and updated set of research priorities that builds on this document will be developed at the 13th World Congress on Endometriosis to be held on May 17-20, 2017, in Vancouver, British Columbia, Canada.
In previous studies, we have shown that menstrual endometrium preferentially adheres to the subepithelial lining of the peritoneum. It remains to be elucidated, however, whether this damage is ...preexisting or inflicted by the menstrual tissue itself. We hypothesized that the menstrual tissue itself damages the peritoneum. To investigate this, the viability of menstrual endometrial tissue in peritoneal fluid (PF) was evaluated and the morphologic changes in the mesothelial cells were studied by in vitro cocultures of menstruum with mesothelial cell monolayers. Menstruum was collected with a menstrual cup. Endometrial tissue was isolated from the menstruum, resuspended in culture medium or in the cell-free fraction of PF and cultured for 24, 48 or 72 h. A 3(4, 5-dimethylthiazolyl-2)-2,5-diphenyl tetrazolium bromide (MTT) assay was performed to obtain a relative measure of viable adhered endometrial cells. Mesothelial cells isolated from human omental tissue were cultured on Matrigel or uncoated plastic. At confluence, overnight cocultures were performed and scanning electron microscopy was used to evaluate the morphologic changes. The viability of endometrial fragments was 84% (n = 36, p < 0.05), 82% (n = 27, not significant) and 104% (n = 14, not significant) when cultured in the cell-free fraction of PF for 24, 48 and 72 h, respectively, when compared to medium with 10% fetal calf serum. Menstrual endometrial fragments or menstrual serum added to and cocultured with mesothelial cells induced severe morphologic alterations of the latter, including retraction, shrinking and gap formation. Similar morphologic changes were observed when mesothelial cells were cocultured with menstrual endometrial fragments in PF or in culture inserts. Incubation with conditioned medium from cultured menstrual endometrium induced similar but less pronounced changes in morphology. In conclusion, menstrual endometrial fragments remain viable in PF in vitro for at least 72 h. Antegradely shed menstruum induces changes in mesothelial cell morphology, including retraction and shrinking with exposure of the underlying surface. These findings suggest that menstruum is harmful to the peritoneal lining. Therefore, by local destruction of the mesothelial layer, menstrual endometrium is able to create sites for adhesion.
Objective: To evaluate whether a menstrual cup is a suitable instrument to collect antegradely shed endometrium for in vitro studies.
Design: A prospective, descriptive, cell biological and ...immunohistochemical study.
Setting: Tertiary care university medical center.
Patient(s): Nine female volunteers with regular cycles.
Intervention(s): Menstrual effluent was collected with a menstrual cup. Experience with the menstrual cup was described. Cytospin specimens, frozen sections, and cultures were prepared from the obtained menstrual tissue.
Main Outcome Measure(s): The acceptability of the menstrual cup. The presence and viability of endometrial tissue was evaluated using immunohistochemical staining and culture outcome.
Result(s): All women except one described the menstrual cup as acceptable. Menstrual effluent contained single cells, clumps of cells, and glandlike structures. After 5 days of culture, the endometrial tissue appeared to be viable. Immunohistochemistry showed positive staining for vimentin in most cytospin specimens, in all cryostat specimens, and in 10 of 17 cultures. Cytokeratin 18 stained most cytospin specimens, all cryostat specimens, and 10 of 17 cultures. Positive staining for BW495/36 was observed in most cytospin specimens, all cryostat specimens, and 11 of 17 cultures.
Conclusion: A menstrual cup is an acceptable instrument to collect antegradely shed menstrual tissue. Menstruum contains viable endometrial tissue that can be used for in vitro studies of endometrium and endometriosis.
To investigate whether the aminoterminal propeptides of type III procollagen are increased in patients with early endometriosis and to demonstrate that the subtle lesion of endometriosis is an active ...stage of the disease.
Aminoterminal propeptide of type III procollagen was determined in serum and peritoneal fluid (PF) of 100 consecutive patients undergoing laparoscopy.
Academisch Ziekenhuis Maastricht, The Netherlands, a tertiary care center.
Aminoterminal propeptide PF levels were significantly higher in women with early lesions of endometriosis compared with levels in two groups of controls, i.e., fertile, cycling, women without the disease (P=0.019) and women on oral contraceptives without the disease (P=0.036). No difference was found in aminoterminal propeptide PF levels when comparing patients with early lesions of endometriosis and patients with unexplained infertility, the third control group. Aminoterminal propeptide PF levels of patients with endometriosis without early lesions were not different from PF levels in controls.
The early lesion is an active stage of endometriosis, invading the extracellular matrix. In women with unexplained infertility active, microscopic endometriosis may be present.
Endometrial wavelike activity was studied by ultrasound, throughout 19 ovulatory cycles in 16 healthy female volunteers. Analysis was focused on the presence of endometrial activity and the wave ...types. Five activity patterns were distinguished, which varied throughout the cycle. Endometrial activity was most striking during the periovulatory phase.
Fertility potential of individual sperm donors Al-Inany, H G; Dunselman, G A; Dumoulin, J C ...
Gynecologic and obstetric investigation,
01/1999, Letnik:
47, Številka:
3
Journal Article
Recenzirano
To study the fertility potential of individual semen donors with ejaculates of optimal as compared to suboptimal quality.
363 semen donations were obtained from 11 donors between January 1993 and ...September 1997. 270 samples were cryopreserved and 1,399 straws obtained from 120 ejaculates were used in 495 insemination cycles.
52 pregnancies were achieved in 128 recipients (40.6%). No significant differences were found between donors of high fecundability and those of low fecundability regarding sperm parameters. The mean donor fecundability index was 10.5%. The optimal and suboptimal samples yielded similar results. Donors with suboptimal semen quality had an increased number of samples rejected after thawing.
Traditional semen analysis parameters do not differentiate high from low fecundability donors. Suboptimal results of the semen analysis should not exclude potential donors from an artificial insemination donor (AID) program.
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