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Biomedical subjects

A F de Goeij

Publications and source records attributed to A F de Goeij.

At least 19 recordsLinked to original sources
PubMed2001

Kirsten ras mutations in patients with colorectal cancer: the 'RASCAL II' study.

Researchers worldwide with information about the Kirsten ras (Ki-ras) tumour genotype and outcome of patients with colorectal cancer were invited to provide that data in a schematized format for inclusion in a collaborative database called RASCAL (The Kirsten ras in-colorectal-cancer collaborative group). Our results from 2721 such patients have been presented previously and for the first time in any common cancer, showed conclusively that different gene mutations have different impacts on outcome, even when the mutations occur at the same site on the genome. To explore the effect of Ki-ras mutations at different stages of colorectal cancer, more patients were recruited to the database, which was reanalysed when information on 4268 patients from 42 centres in 21 countries had been entered. After predetermined exclusion criteria were applied, data on 3439 patients were entered into a multivariate analysis. This found that of the 12 possible mutations on codons 12 and 13 of Kirsten ras, only one mutation on codon 12, glycine to valine, found in 8.6% of all patients, had a statistically significant impact on failure-free survival (P = 0.004, HR 1.3) and overall survival (P = 0.008, HR 1.29). This mutation appeared to have a greater impact on outcome in Dukes' C cancers (failure-free survival, P = 0.008, HR 1.5; overall survival P = 0.02, HR 1.45) than in Dukes' B tumours (failure-free survival, P = 0.46, HR 1.12; overall survival P = 0.36, HR 1.15). Ki-ras mutations may occur early in the development of pre-cancerous adenomas in the colon and rectum. However, this collaborative study suggests that not only is the presence of a codon 12 glycine to valine mutation important for cancer progression but also that it may predispose to more aggressive biological behaviour in patients with advanced colorectal cancer.

Adolescent↗
PubMed2001

Tumor necrosis factor-alpha but not interleukin-1 beta or interleukin-8 concentrations correlate with angiogenic activity of peritoneal fluid from patients with minimal to mild endometriosis.

OBJECTIVE: To assess the angiogenic activity of peritoneal fluid in women with minimal to mild endometriosis and to investigate the relationship between this activity and the concentration of macrophage-derived angiogenic factors and clinical variables, such as phase of menstrual cycle, type of lesion, and revised American Society for Reproductive Medicine classification. DESIGN: In vivo bioassay. SETTING: Tertiary-care university medical center. PATIENT(S): Fifty-two female volunteers with laparoscopic findings indicating minimal to mild endometriosis. INTERVENTION(S): Peritoneal fluid was collected at the start of laparoscopy. A standard amount of peritoneal fluid was applied to a chick embryo chorioallantoic membrane assay. MAIN OUTCOME MEASURE(S): Angiogenic response was assessed by determining the vascular density index. RESULT(S): 85% of the peritoneal fluid samples induced angiogenesis in the chick embryo chorioallantoic membrane bioassay. Tumor necrosis factor-alpha and total protein were significantly related to the vascular density index, whereas interleukin-1beta, interleukin-8, and clinical variables appeared to not affect the angiogenic response. CONCLUSION(S): The results confirms previous findings of peritoneal fluid angiogenic activity in women with minimal to mild endometriosis and indicate involvement of tumor necrosis factor-alpha.

Adult↗
PubMed2001

The Mesothelium, Teflon or Velcro? Mesothelium in endometriosis pathogenesis.

Sampson's transplantation theory for the pathogenesis of peritoneal endometriosis is widely accepted. The events that take place, however, on the cellular and subcellular level during the transition of endometrial tissue in the abdominal cavity into peritoneal endometriosis remain controversial. The mesothelium plays a central role in the debate on this subject. The interaction between endometrium and peritoneum has been studied in an in-vitro model using amnion, peritoneum and mesothelial cells in culture on the one hand and cyclic and menstrual endometrium on the other hand. The results of these studies indicate that (i) an intact mesothelial lining prevents adhesion of shed endometrial tissue, (ii) shed endometrial tissue adheres to the peritoneal extracellular matrix and (iii) menstrual effluent creates its own adhesion sites by damaging the mesothelial lining thus exposing the extracellular matrix. Therefore we conclude that the mesothelium has the properties of Teflon, while the extracellular matrix resembles Velcro.

Endometriosis↗
PubMed2001

Development of endometriosis-like lesions after transplantation of human endometrial fragments onto the chick embryo chorioallantoic membrane.

The chick embryo chorioallantoic membrane (CAM) bioassay was used to investigate the early pathogenesis of endometriosis. Endometrial fragments were explanted onto the CAM. The grafts including the surrounding CAM were excised at 24, 48 or 72 h after explantation, fixed and embedded in paraffin. Immunohistochemical analysis was used to distinguish endometrial cells. To identify cells of human origin, in-situ hybridization was performed using a probe specific for human chromosome 1. After 24 h, direct contact between endometrial stromal as well as epithelial cells and the mesenchymal layer of the CAM was observed. Invasion of both stromal cells and intact endometrial glands into the mesenchymal layer was observed after 48 h. At 72 h, endometriosis-like lesions were observed in the mesenchymal layer. Positive staining with antibodies to vimentin and pan-cytokeratin was observed in the invading cells as well as in the lesions. In the lesions these positively stained cells showed in-situ hybridization signals for human chromosome 1, confirming their human origin. In conclusion, after 3 days of incubation, endometriosis-like lesions consisting of human endometrial glands and stromal cells were found in the mesenchymal layer of the CAM. These lesions apparently resulted from the invasion of intact human epithelial structures and stromal cells.

Allantois↗
PubMed2001

Endometrial angiogenesis throughout the human menstrual cycle.

BACKGROUND: The timing and mechanisms of new blood vessel formation in the endometrium during the menstrual cycle are still largely unknown. In the present study we used the chick embryo chorioallantoic membrane (CAM) as an in-vivo assay for angiogenesis to assess the angiogenic potential of endometrium obtained at different stages of the menstrual cycle. METHODS: Endometrial fragments were explanted onto the CAM and, after 4 days of incubation, slides of the treated area were taken in ovo through a microscope for computerized image analysis. The vascular density index (VDI), a stereological estimate of vessel number and length, was obtained by counting the intersections of vessels with five concentric circles of a circular grid superimposed on the computerized image. RESULTS: We demonstrated that human endometrium has angiogenic potential throughout the menstrual cycle. Furthermore, there was a significant difference in angiogenic response between the stages of the menstrual cycle (P = 0.01). The VDIs of the early proliferative, early and late secretory stage were significantly higher than the VDI of the late proliferative phase. CONCLUSIONS: Elongation of existing vessels during the early proliferative phase as well as growth and coiling of the spiral vessels during the secretory phase may demand far higher angiogenic activity than outgrowth and maintenance of vessels during the late proliferative phase.

Allantois↗
PubMed2001

Molecular genetic alterations in hamartomatous polyps and carcinomas of patients with Peutz-Jeghers syndrome.

AIM: To investigate whether mutations in the STK11/LKB1 gene and genes implicated in the colorectal adenoma-carcinoma sequence are involved in Peutz-Jeghers syndrome (PJS) related tumorigenesis. METHODS: Thirty nine polyps and five carcinomas from 17 patients (from 13 families) with PJS were analysed for loss of heterozygosity (LOH) at 19p13.3 (STK11/LKB1 gene locus), 5q21 (APC gene locus), 18q21-22 (Smad4 and Smad2 gene locus), and 17p13 (p53 gene locus), and evaluated for immunohistochemical staining of p53. In addition, mutational analysis of K-ras codon 12, APC, and p53 and immunohistochemistry for Smad4 expression were performed on all carcinomas. RESULTS: LOH at 19p was seen in 15 of the 39 polyps and in all carcinomas (n = 5). Interestingly, six of the seven polyps from patients with cancer had LOH, compared with nine of the 31 polyps from the remaining patients (p = 0.01). In one polyp from a patient without a germline STK11/LKB1 mutation, no LOH at 19p or at three alternative PJS candidate loci (19q, 6p, and 6q) was found. No LOH at 5q was observed. However, mutational analysis revealed an APC mutation in four of the five carcinomas. LOH at 17p was not seen in polyps or carcinomas; immunohistochemistry showed expression of p53 in one carcinoma and focal expression in three polyps. At subsequent sequence analysis, no p53 mutation was found. One carcinoma had an activating K-ras codon 12 mutation and another carcinoma showed 18q LOH; however, no loss of Smad4 expression was seen. CONCLUSIONS: These results provide further evidence that STK11/LKB1 acts as a tumour suppressor gene, and may be involved in the early stages of PJS tumorigenesis. Further research is needed to see whether LOH in PJS polyps could be used as a biomarker to predict cancer. Differences in molecular genetic alterations noted between the adenoma-carcinoma sequence and PJS related tumours suggest the presence of a distinct pathway of carcinogenesis.

AMP-Activated Protein Kinase Kinases↗
PubMed2000

Matrix metalloproteinases and their tissue inhibitors in antegradely shed menstruum and peritoneal fluid.

OBJECTIVE: To investigate the expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in antegradely shed menstruum and peritoneal fluid. DESIGN: A cell biological and immunohistochemical study. SETTING: Tertiary care university medical center. INTERVENTION(S): Immunohistochemistry was performed on cryostat sections and cultures of menstrual endometrium. Zymography was used to characterize MMP activity in peritoneal fluid, in menstrual serum, and in conditioned medium. Western blot analysis was used to further identify the MMPs in these fluids. MAIN OUTCOME MEASURE(S): Staining of MMPs and TIMPs in cryostat sections and cultures and MMP expression and activity in peritoneal fluid and menstrual blood serum. RESULT(S): Strong staining for MMP-1 and MMP-3 was observed in stroma and for MMP-7 in epithelium. Matrix metalloproteinase-2 and MMP-9 were weakly expressed in stroma. Both TIMP-1 and TIMP-2 were expressed in menstrual endometrium. Menstrual serum showed a pattern of MMP activity on zymography different from peritoneal fluid. Western blot analysis showed the presence of MMP-7 and MMP-9 in menstrual serum. CONCLUSION(S): Antegradely shed menstrual endometrium expresses several MMPs and TIMPs, even after culturing for 24 hours. MMP activity in menstrual serum is different from and more intense than MMP activity in peritoneal fluid. These enzymes may be involved in the early invasion of menstrual endometrium into the extracellular matrix of the peritoneum.

Ascitic Fluid↗
PubMed2000

Morphological changes in mesothelial cells induced by shed menstrual endometrium in vitro are not primarily due to apoptosis or necrosis.

In a previous study on the pathogenesis of endometriosis, we observed that constituents of menstrual effluent induce morphological alterations in human mesothelial cells. In this study, we investigated whether these alterations were associated with apoptosis or necrosis or were the result of cellular remodelling. After overnight incubation of confluent monolayers of human omental mesothelial cells (HOMEC) with conditioned media prepared from menstrual effluent shed anterogradely, severe alterations in morphology were observed. Typical polygonal mesothelial cell cultures at confluency acquired elongated spindle morphology, resulting in gaps between the cells. In contrast, mesothelial cells from the control groups receiving culture medium only, retained a normal morphology. Immunofluorescence staining revealed that cytokeratin, vimentin and actin filaments were still present, homogeneously distributed in the cell cytoplasm following changes in morphology. To evaluate whether the morphological alterations were associated with apoptosis and/or necrosis, the cells were stained with the M30 CytoDeath antibody or annexin V with propidium iodide and analysed using flow cytometry. The results showed that only a small percentage (1-7%) of the affected HOMEC were undergoing apoptosis or necrosis. We conclude that the profoundly altered morphology of HOMEC is a result of cellular remodelling and that the role of apoptosis and necrosis is negligible. Soluble paracrine factors released by cells isolated from menstrual effluent shed anterogradely may induce a reorganization of the cytoskeleton. As a result, the underlying basement membrane will be exposed and the mesothelium may no longer prevent implantation of endometrium shed retrogradely into the peritoneum, thus facilitating the development of endometriosis.

Annexin A5↗
PubMed2000

Adhesion of menstrual endometrium to extracellular matrix: the possible role of integrin alpha(6)beta(1) and laminin interaction.

Previous in-vitro studies have shown that the endometrium preferentially adheres to the extracellular matrix (ECM) of the amnion and peritoneum. This interaction probably involves adhesion molecules, e.g. integrins. We evaluated the expression of integrins in naturally shed menstrual endometrium and the adhesion pattern of this tissue to different components of the ECM. To identify integrins and matrix components involved, blocking studies were performed. Most of the 15 menstrual tissue samples showed positive staining for each of the integrins investigated, except alpha(4)beta(1). Compared with binding to collagen IV, which was set at 100%, adhesion to collagen I was 93% (not significant), to fibronectin 87% (P < 0.05), and to laminin 74% (P < 0.05). Scanning electron micoscopy showed that endometrium adhered to laminin but hardly spread, whereas spreading was observed when layered on the other coatings. Compared with the control (which was set at 100%), incubation with 4B4, a monoclonal antibody against the integrin beta(1) subunit, showed a significant reduction of adhesion (to approximately 50%; P < 0.05) when layered on laminin and a smaller reduction (to 82-86%; P < 0.05) when layered on the other three coatings. Incubation with antibody GOH3 against integrin alpha(6)beta(1) resulted in a similar reduction in adhesion to laminin. Incubation with an RGD peptide significantly reduced adhesion (to 84%; P < 0.05) when plated on fibronectin. In conclusion, antegradely shed menstrual endometrium expresses various integrins. It shows preferential attachment to collagen IV and collagen I, when compared with fibronectin and laminin. Blockage of the integrin beta(1) subunit resulted in greatest disruption to adhesion when layered on laminin, implying that the interaction was mediated by the alpha(6)beta(1) integrin. Since this adhesion was not completely blocked, other mechanisms are likely to be involved.

Adult↗
PubMed2000

Menstruum induces changes in mesothelial cell morphology.

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.

Adipocytes↗
PubMed1999

Adhesion of human endometrial fragments to peritoneum in vitro.

OBJECTIVE: To evaluate the adhesion of endometrial fragments obtained during the proliferative phase of the menstrual cycle to fresh human peritoneum obtained during abdominal surgery. DESIGN: A prospective, descriptive, morphologic and cell biologic study. SETTING: Tertiary care university medical center. PATIENT(S): Six female volunteers. INTERVENTION(S): After endometrial biopsies performed during diagnostic laparoscopy, endometrial fragments were generated by enzymatic digestion and mechanical separation. Peritoneum was obtained during abdominal operations for benign indications. MAIN OUTCOME MEASURE(S): Adhesion of endometrial fragments was studied by histologic examination and scanning and transmission electron microscopy. RESULT(S): After incubation, the mesothelium was intact in some areas, whereas in other areas mesothelial cells were damaged or absent. Adhesion of endometrial fragments was observed only at locations where the basement membrane was exposed. In areas largely denuded of mesothelial cells, endometrial fragments spread over the basement membrane to form monolayers. CONCLUSION(S): Human peritoneum is suitable for studying the adhesion of endometrial fragments. Intact mesothelium prevents the adhesion of endometrial fragments, suggesting that trauma to the mesothelial lining is a prerequisite for endometrial cell adhesion.

Basement Membrane↗
PubMed1999

Adhesion of shed menstrual tissue in an in-vitro model using amnion and peritoneum: a light and electron microscopic study.

We have investigated the adhesion of endometrial tissue isolated from antegradely shed menstrual effluent to amnion and peritoneum. This endometrial tissue was cultured overnight on either side of intact and stripped amnion and on the mesothelial side of peritoneum. Light and electron microscopy were applied to evaluate adhesion. With light microscopy adhesion of endometrial fragments to stripped membranes was observed in nine out of 12 specimens and in 12 out of 13 specimens when layered on the extracellular matrix side of amnion. Adhesion when layered on the epithelial side was seen in only four out of 13 specimens. However, when using scanning electron microscopy adhesion of menstrual endometrial tissue could be visualized in all samples. Numerous adhering fragments were seen when layered on the extracellular matrix side of untreated amnion. On several occasions not only adhesion but also spreading of cells was observed. When layered on the epithelial side of untreated amnion or peritoneum, adhesion was exclusively seen at locations where the epithelium was damaged or absent. These findings were confirmed by transmission electron microscopy. These observations indicate that endometrial tissue isolated from antegradely shed menstrual effluent preferentially adheres to subepithelial structures of amnion and peritoneum. The lack of adhesion to epithelial cells suggests that an intact mesothelial lining prevents adhesion of menstrual endometrial tissue.

Amnion↗
PubMed1999

The chick embryo chorioallantoic membrane as a model to investigate the angiogenic properties of human endometrium.

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.

Allantois↗
PubMed1998

Adhesion of human endometrium to the epithelial lining and extracellular matrix of amnion in vitro: an electron microscopic study.

One of the first steps in the pathogenesis of endometriosis is the attachment of the endometrium to the peritoneal lining. Since the peritoneum is extremely fragile and hard to obtain, amnion has been used as an in-vitro model to study adhesion. Scanning and transmission electron microscopy was applied to evaluate the adhesion of endometrial cells isolated in the proliferative and secretory phases of the menstrual cycle. Endometrial fragments obtained in either phase of the cycle were able to adhere to the extracellular matrix of the amnion. Fragments from proliferative phase endometrium showed active spreading and growth over the matrix surface, whereas fragments from secretory phase endometrium did not. Fragments from proliferative as well as secretory phase endometrium were able to adhere to the epithelial side of the amnion, but only at locations where the amniotic epithelium was damaged or partly absent. These observations indicate that the basement membrane and extracellular matrix provide a suitable substrate for endometrial cell attachment and growth and that endometrial cell adhesion occurs preferentially to subepithelial structures, whereas an intact epithelium prevents the adhesion of endometrial fragments to the amnion.

Amnion↗
PubMed1998

Amniotic membrane as an in vitro model for endometrium-extracellular matrix interactions.

The aim of the study was to develop an in vitro model to study the interaction between endometrial cells and extracellular matrix (ECM) and to evaluate the expression of cell adhesion molecules in endometrial cells and tissue fragments under in vitro conditions. Endometrial biopsies were collected from 32 patients. Samples were either digested using collagenase type I, or dissected mechanically. Adhesion of isolated cells and tissue fragments to stripped amniotic membranes and to coverslips coated with ECM components was studied. The expression of beta1 integrins and cadherins was assessed using immunohistochemistry. Collagenase digestion of endometrial biopsies yielded viable single cells. These cells did not adhere to either side of stripped amniotic membranes, and did not show expression of the cell adhesion molecules. In contrast, mechanically fragmented endometrium samples adhered to both sides of stripped amniotic membranes and showed immunohistochemical expression of E- and P-cadherin and integrin subunits alpha2, alpha3, alpha4, alpha5 and alpha6. Amniotic membranes, after stripping of epithelial lining, are suitable to study interactions between endometrial tissue and ECM in functional and structural studies. Endometrial cells after collagenase type I digestion do not adhere to stripped amniotic membrane and have lost expression of beta1 integrins and E- and P-cadherin.

Amnion↗
PubMed1997

Evaluation of a menstrual cup to collect shed endometrium for in vitro studies.

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 in 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.

Adult↗
PubMed1996

Endometrial cell adhesion in an in vitro model using intact amniotic membranes.

OBJECTIVE: To study adhesion between endometrial tissue and peritoneum using intact amniotic membranes as an in vitro model. DESIGN: A cell biologic and immunohistochemical study. SETTING: Tertiary-care university medical center. PATIENTS: Ten patients with regular and ovulatory cycles. INTERVENTIONS: 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. MAIN OUTCOME MEASURES: 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. RESULTS: 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. CONCLUSIONS: An intact epithelial lining prevents adhesion of endometrial fragments to the ECM of amniotic membranes in vitro. An intact epithelium could be important defense mechanism in preventing initial adhesion of retrogradely shed endometrium fragments to peritoneum.

Amnion↗