Growth restricting effects of a single course of antenatal betamethasone treatment and the role of human placental lactogen.

UNLABELLED: Betamethasone (BET) is a widely used treatment for women who are at high risk of preterm delivery. In sheep, BET-induced growth restriction was found to be associated with reduced placenta lactogen (PL), a key regulator of fetal growth. We therefore hypothesized that also in humans a single course of BET administration is associated with a reduction of PL, associated with a deceleration in fetal growth. OBJECTIVE: To investigate effects of a single course of antenatal BET in humans on birth weight and PL. METHODS: Women exposed to BET (2 × 12 mg; n = 44) with normally grown fetuses between 23 + 5 and 34 + 0 wks (weeks + days of gestation) who delivered between 23 + 5 to 42 + 0 wks were compared to gestational age-matched controls (n = 49). Maternal gestational blood samples were obtained before, during and after BET treatment and at the time of birth. MAIN OUTCOME MEASURES: BET effects on fetal anthropometrics, placental morphometry and placental PL-protein and maternal plasma levels. RESULTS: The mean duration of days between BET administration and birth was 52 days. BET treatment was associated with decreased birth weight (-18.2%), head circumference (-8.6%), body length (-6.0%), and placental width (-5.5%), as compared to controls. These changes were irrespective of possible maternal confounders (gestational age at birth, maternal age, maternal BMI gain during pregnancy, smoking etc.). However, neither PL-plasma levels within 48 h after BET treatment nor placental PL-protein levels and maternal plasma levels at birth were changed after BET treatment. In central regions of the placenta, BET treatment increased the circumference of syncytiotrophoblast nuclei by +4.7% and nucleus surface area by +9.4% compared to controls, but these changes were not related to placental PL-protein or maternal PL-plasma levels at birth. CONCLUSION: A single course of BET treatment was accompanied with reduced fetal growth, but this growth restricting effect was not associated with altered placental or maternal plasma PL levels. Altered expression of PL appears not to be causal for BET-induced fetal growth restriction in the human.

Association of placental inflammation with fetomaternal hemorrhage and loss of placental mucin-1.

BACKGROUND: Fetomaternal hemorrhage (FMH) poses an immediate risk to the fetus and, in case of Rhesus-immunization, to future pregnancies. Given that altered endothelial permeability is part of the pathophysiology of inflammation, in this study we investigated whether placental inflammatory processes like chorioamnionitis (ChoA) or preeclampsia (PE) lead to increased rates of FMH compared to the established risk factor of placenta previa (PP). Putative accompanying markers of trophoblastic damage were also explored. METHODS: 40 patients (14 PE; 6 ChoA; 9 PP; 11 normal controls) were evaluated for FMH using a flowcytometric test kit, which is able to quantify FMH of 0.06% fetal cells. Placental tissue samples were immunostained for human placental lactogen (hPL), human chorionic gonadotropin (hCG), and mucin-1 (MUC1). MUC1 was evaluated as a potential serum marker of FMH. RESULTS: Patients with ChoA had a mean calculated FMH volume of 29 ml, compared to 4 ml in PE and 1 ml in PP and controls. MUC1 staining was reduced in PE and ChoA placenta samples, while elevated MUC1 serum concentration correlated positively with FMH. CONCLUSION: Diseases of placental inflammation are associated with FMH. Placental MUC1 staining is reduced and serum concentrations are increased in cases of FMH.

A placental chorionic villous mesenchymal core cellular origin for infantile haemangioma.

AIMS: To investigate the expression of the placental cell-specific associated proteins in infantile haemangioma (IH). METHODS: Immunohistochemical staining was used to investigate the expression of human chorionic gonadotrophin (hCG), human placental lactogen (hPL), human leucocyte antigen-G (HLA-G), cytokeratin 7 (CK7) and smooth muscle actin in paraffin-embedded sections of proliferating and involuted IHs. RESULTS: The proteins hCG and hPL were expressed by the endothelium but not the pericyte layer of proliferating IH, but these proteins were not detected in involuted lesions. There was no expression of CK7 and HLA-G in IH. CONCLUSIONS: The expression of hCG and hPL, but not CK7 or HLA-G, by the endothelium of proliferating IH supports a placental chorionic villous mesenchymal core cellular origin for IH rather than a trophoblast origin.

Proteomic analysis of human serum for finding pathogenic factors and potential biomarkers in preeclampsia.

OBJECTIVE(S): To apply a novel proteomic method to discover potential pathogenic factors and biomarkers of preeclampsia. STUDY DESIGN: Sera from five patients complicated with preeclampsia and five healthy pregnant controls were separately pooled. Each pool was treated with peptide ligand library beads (PLLBs) to remove high abundance proteins by affinity and thus enrich low abundance proteins. The proteins from the eluate were analyzed by a combination of 1D Gel-LC-MS/MS. Protein expression levels were quantified using spectral counts and the extracted ion current. RESULTS: 1172 unique proteins in preeclampsia and 1149 in healthy controls were identified in the present study. 51 proteins were differentially expressed between preeclampsia and healthy pregnant women including chorionic somatommammptropin hormone (CSH) and fibulin-1. 31 proteins identified were up-regulated and 20 were down-regulated. CONCLUSIONS: The results demonstrate that peptide ligand library combining with 1D gel-LC-MS/MS analysis is an efficient method to identify differentially expressed proteins in sera and two biological processes of complement and coagulation activations and lipid metabolism were involved in the pathogenesis of preeclampsia.

Specificity and sensitivity of differentiation antigens in superficial soft tissue tumors: comparison of SMA, calponin, H-caldesmon, C-kit, PLAP and HPL.

We examined the expression pattern of smooth muscle actin (SMA), h-caldesmon (HCD), calponin (CALP), placental alkaline phosphatase (PLAP) and human placental lactogen (HPL) in benign and malignant spindle cell superficial soft tissue tumors in order to determine the role of these markers in differential diagnosis. Archival tissue from 38 patients with superficial smooth muscle cell and so-called fibrohistiocytic tumors (8 benign fibrous histiocytomas (BFHs), 6 dermatofibrosarcoma protuberans (DFPT), 9 malignant fibrous histiocytomas (MFHs), 9 leiomyomas (LMs) and 6 leiomyosarcomas (LMSs)) were immunostained with antibodies against SMA, HCD, CALP, PLAP and HPL. smooth muscle cell (SMC) tumors showed significantly high immunopositivity for HCD than that of so-called fibrohistiocytic tumors (p is less than or equal to 0.05) but 1/3 of DFPT and MFH cases and half of BFH cases also showed HCD immunopositivity; thus, this difference is debatable and not highly discriminative as expected. All tumor groups showed 100% immunopositivity for CALP. SMC tumors displayed significantly stronger and more widespread immunostaining pattern for PLAP than so-called fibrohistiocytic tumors (p < 0.05). Superficial soft tissue tumors did not express c-kit. In conclusion, HCD and PLAP can be used as ancillary immunomarkers in differential diagnosis of SMC tumors (Tab. 2, Fig. 7, Ref. 37).

Severe spontaneous acute tumor lysis syndrome and hypoglycemia in patient with germ cell tumor.

Tumor lysis syndrome has been observed in patients with bulky, treatment-sensitive tumors, in particular hematological malignancies, especially after medical treatment (chemotherapy, corticosteroids, radiation, hormonal agents, and biological response modifiers). Tumor lysis syndrome has been observed also in solid malignancies and it very rarely occurs spontaneously. Tumor lysis syndrome-associated metabolic abnormalities include hyperuricemia, hyperphosphatemia, hyperkalemia, hypocalcemia and uremia. Severe hypoglycemia is another rare metabolic disorder, uncommonly associated with solid malignancies. The case described here is peculiar for the abrupt onset of these two rare conditions in a patient with a metastatic germ cell tumor.

Primary choriocarcinoma of the liver: a clinicopathological study of five cases in males.

Primary choriocarcinomas of the liver are rare. Previous reported cases were mostly in infants with only rare adult cases. Here, we presented five adult cases. The patients were all males, with an average age of 41.6 years (from 36 to 48 years). Clinical presentations included right upper abdominal pain or abdominal distension. All patients presented with a large hepatic mass on ultrasonography that measured 11 cm on average in the greatest diameter. Elevated serum human chorionic gonadotropin (HCG) levels were noted in all cases. At presentation, the tumor was confined to the liver in two patients and therefore surgically resected. The other three patients presented with extrahepatic metastases on imaging study and therefore only received chemotherapy. All five patients died from the tumor within 2 to 8 months. Autopsy was performed for all five cases. The autopsy confirmed that the choriocarcinoma was confined to the liver in two surgically resected cases. The other three patients had metastatic choriocarcinoma in the lung (two patients), peritoneum (one patient), adrenal glands (one patient), and brain (one patient). None of the patients had any evidence of a testicular tumor or scar after examination of the entirely submitted testes. No tumor was observed in central nervous system, mediastinum, or other organs other than described above. Grossly, the primary tumors were large, soft, hemorrhagic, and with foci of necrosis. Histologically, the tumors were composed of mononucleated trophoblastic cells with round nuclei, clear cytoplasm, and prominent nucleoli admixed with large, multinucleated syncytiotrophoblastic cells. Immunohistochemically, tumor cells were strongly positive for keratin, HCG, and focally positive for human placental lactogen. Ki-67 proliferation index was high (mean 75%) in the mononucleated trophoblastic cells. Our series is the largest one to document primary hepatic choriocarcinoma in adults. Although these tumors are rare, they behave in a very aggressive fashion.

P57kip2 immunohistochemical expression and ultrastructural findings of gestational trophoblastic disease and related disorders.

Gestational trophoblastic disease (GTD) is a unique spectrum of diseases ranging from complete hydatidiform mole (CHM), partial hydatidiform mole (PHM), and invasive mole (IM) to choriocarcinoma (CC). Placental site trophoblastic tumor (PSTT) and epithelioid trophoblastic tumor (ETT) have been classified as related disorders. Mesenchymal dysplasia (MD) may be misdiagnosed as PHM; however, it is said to have a quite different histogenesis from PHM. P57kip2 is the protein product of a paternally imprinted or maternal gene that inhibits cyclin-dependent kinases (CDK), thus serving to inhibit cell proliferation and to suppress tumor growth. Its lack of expression in trophoblastic disease plays a role in its abnormal proliferation and differentiation. In this study, P57kip2 immunostaining was absent in the trophoblastic layers of CHM and was positive in the trophoblast layer of nonmolar villi and MD. Ultrastructure of complete molar cystic villi showed tree-like branching of microvillous processes and intracytoplasmic lacunae without capillaries in the stroma, whereas MD contained many newly formed blood vessels and collagen. Also, large lacunae with microvilli and polymorphic nuclei of syncytiotrophoblast cells with well-developed organelles were observed in IM. Lung ETT following CHM and normal deliveries showed two types of large mononuclear cells and binuclear cells with abundant organelles and bundles of intermediate-type filaments in the stroma.

In vitro differentiation of villous trophoblasts from pregnancies complicated by intrauterine growth restriction with and without pre-eclampsia.

Highly purified (>99.99%) primary villous cytotrophoblasts from uncomplicated pregnancies and pregnancies complicated with intrauterine growth restriction (IUGR) alone, IUGR with pre-eclampsia (IUGR-PE) and PE alone were cultured for 5days and the extent of differentiation into syncytiotrophoblasts measured in terms of syncytialisation and secretion of chorionic gonadotropin (hCG) and placental lactogen (hPL). Three separate phenotypes were observed: (1) normal and IUGR-PE cells showed low syncytialisation and secretion of hCG and hPL, (2) IUGR cells showed the highest levels of syncytialisation and secretion and (3) PE cells showed high syncytialisation but low secretion. These results strongly suggest IUGR, IUGR-PE and PE to be distinct conditions in which villous cytotrophoblasts are either exposed to different environments or are genetically different.

Relative Concentrations of Placental Lactogen II and PRL-Like Protein-A in Stressed Rats Placenta / Concentraciones del Lactógeno Placentario II y la Proteína A Ligada a Prolactina en Placentas de Ratas Estresadas

The chronic stress induces functional adaptations in the hypothalamo-pituitary- adrenocortical (HPA) and in the sympathetic-medullary-adrenal axis (SAM). Both axis are considered vital regulators of the homeostasis in vertebrates (Seyle, 1936; Ostrandrer et al, 2006. On the other hand, the placenta provides highly specialized functions during gestation that are critical for the normal development of the embryo/fetus (Soares et al., 1991). We hypothesized that the chronic immobilization (IMO) stress in pregnancy rats produces alterations in prolactin concentrations in placental tissue and also changes in the response of SAM axis. Chronic stress by IMO was applied on days 12, 17 and 21 of pregnancy rats. Relative concentrations and localization of placental lactogen-II (PL-II) and the PRL- like protein A (PLP-A) in chorioalantoic placenta were estimated by Immunoblotting and Immunocytochemical analysis. The levels of catecholamines metabolite, acid 3-metoxi 4-hidroximandélico (VMA), were analyzed in stressed rats urines on 6,12,17,21 days of pregnancy, by HPLC, in order to determine the response of SAM axis. During the days of the pregnancy studied, chronic stress did not induce any changes neither in the localization nor in placental concentrations of PL-II and PLP-A. The VMA values in stressed mothers urines increased on the day 6 respecting the control ones at the same time of pregnancy. VMA values in stressed rats at 21 days of pregnancy are smaller than the respective controls. We conclude that the chronic stressed mothers activated the SAM axis at the beginning of pregnancy and then they diminished the metabolites catecholamines that were interpreted as a stress adaptation coincident with normal concentrations of both placentary prolactines at this stage of the pregnancy.

El estrés crónico induce adaptaciones funcionales en los ejes hipotálamo-pituitario-adrenal (UPA) y en el simpático médulo adrenal (SAM). Ambos ejes son considerados reguladores vitales de la homeostasis en los vertebrados (Seyle, 1936; Ostrandrereí al., 2006). Por otro lado, el desarrollo y crecimiento fetal de los mamíferos dependen en gran medida del buen funcionamiento de la placenta (Soares, 1991). Nosotros hipotetizamos que el estrés crónico por inmovilización (IMO) aplicado a las ratas gestantes produce alteraciones en las concentraciones de las prolactinas en el tejido placentario y cambios en la respuesta del eje SAM. Se le aplicó estrés crónico por IMO a las hembras en los días 12, 17 y 21 de la preñez y se analizó por inmunocitoquímica e inmunoblotting la localización y concentraciones del lactógeno placentario dos (PL-II) y la proteína A ligada a la prolactina (PLP-A) en la placenta. Se analizaron por HPLC, en las orinas de ratas preñadas (6,12,17,21 días), los niveles del metabolito de las catecolaminas, (ácido 3-metoxi 4-hidroximandélico) (VMA), a fin de determinar la respuesta del eje SAM al tratamiento. El estrés crónico no indujo cambios tanto en la localización como en las concentraciones de PL-II y PLP-A en las placentas en los días de la preñez estudiados. Los valores de VMA en las orinas de las madres estresadas se incrementaron en el día 6 con respecto al control del mismo tiempo de preñez. Mientras que a los 21 días los valores de VMA de las ratas estresadas son menores que los controles respectivos. Concluimos que en las madres estresadas crónicamente, no se alteraron las concentraciones de ambas prolactinas placentarias. En cambio se activó el eje SAM al comienzo de la preñez ante el primer estímulo estresante y luego una reducción de la respuesta del eje ante el estrés crónico, a medida que avanza la preñez.

Circulating cell-free fetal messenger RNA levels after fetoscopic interventions of complicated pregnancies.

OBJECTIVE: The aim of this study was to examine fetal gene expression in maternal plasma after fetoscopic intervention for twin-twin transfusion syndrome or congenital diaphragmatic hernia. STUDY DESIGN: Twelve women with pregnancies that were complicated by twin-twin transfusion syndrome and 10 women carrying fetuses with congenital diaphragmatic hernia were sampled before and sequentially after treatment. Levels of glyceraldehyde-3-phosphate dehydrogenase, human placental lactogen, and gamma globin messenger RNA were measured by real-time reverse transcriptase polymerase chain reaction amplification. RESULTS: At all time points, glyceraldehyde-3-phosphate dehydrogenase messenger RNA levels were higher in the congenital diaphragmatic hernia cases than in the twin-twin transfusion syndrome cases (P < .05), but during the immediate postoperative observation period, there were no significant changes in glyceraldehyde-3-phosphate dehydrogenase, human placental lactogen, or gamma globin messenger RNA levels in individual patients or patients who were grouped by procedure. CONCLUSION: Fetoscopic intervention of complicated pregnancies does not affect circulating fetal messenger RNA levels, which is in contrast to earlier observations that circulating fetal DNA levels increase after laser ablation for twin-twin transfusion syndrome. Plasma glyceraldehyde-3-phosphate dehydrogenase messenger RNA levels could be a potential novel biomarker for fetal trauma.

Hypoxia inhibits differentiation of lineage-specific Rcho-1 trophoblast giant cells.

Defects in placental development lead to pregnancies at risk for miscarriage and intrauterine growth retardation and are associated with preeclampsia, a leading cause of maternal death and premature birth. In preeclampsia, impaired placental formation has been associated with alterations in a specific trophoblast lineage, the invasive trophoblast cells. In this study, an RT-PCR Trophoblast Gene Expression Profile previously developed by our laboratory was utilized to examine the lineage-specific gene expression of the rat Rcho-1 trophoblast cell line. Our results demonstrated that Rcho-1 cells represent an isolated, trophoblast population committed to the giant cell lineage. RT-PCR analysis revealed that undifferentiated Rcho-1 cells expressed trophoblast stem cell marker, Id2, and trophoblast giant cell markers. On differentiation, Rcho-1 cells downregulated Id2 and upregulated Csh1, a marker of the trophoblast giant cell lineage. Neither undifferentiated nor differentiated Rcho-1 cells expressed spongiotrophoblast marker Tpbpa or labyrinthine markers Esx1 and Tec. Differentiating Rcho-1 cells in hypoxia did not alter the expression of lineage-specific markers; however, hypoxia did inhibit the downregulation of the trophoblast stem cell marker Id2. Differentiation in hypoxia also blocked the induction of CSH1 protein. In addition, hypoxia inhibited stress fiber formation and abolished the induction of palladin, a protein associated with stress fiber formation and focal adhesions. Thus, Rcho-1 cells can be maintained as a proliferative, lineage-specific cell line that is committed to the trophoblast giant cell lineage on differentiation in both normoxic and hypoxic conditions; however, hypoxia does inhibit aspects of trophoblast giant cell differentiation at the molecular, morphological, and functional levels.

Expression profile and protein levels of placental products as indirect measures of placental function in in vitro-derived bovine pregnancies.

Bovine conceptus development and its association with placental proteins present in maternal, foetal and neonatal plasma and foetal (amniotic and allantoic) fluids were investigated in in vivo- and in vitro-produced (IVP) concepti and newborn calves. Females were superovulated to obtain control embryos, whereas IVP embryos were derived from established in vitro procedures. Pregnant animals were slaughtered on days 90 or 180 of gestation or allowed to develop to term for the assessment of physical traits. Foetal, maternal and neonatal blood and foetal fluids were collected for the determination of bovine placental lactogen (bPL) and bovine pregnancy-specific protein B (bPSPB) concentrations. Placental transcripts for bPL and bPSPB, determined by quantitative RT-PCR, were elevated in IVP placentomes. No major physical differences were observed between groups on day 90, but concentrations of bPL and bPSPB were higher in foetal plasma and allantoic fluid of IVP concepti in day 180 pregnancies, which were correlated with larger uterine and conceptus traits. Maternal concentrations of bPL in IVP pregnancies were lower than controls during the last 8 weeks of gestation, to become similar as parturition approached. Newborn IVP calves and foetal membranes were larger and displayed higher concentrations of plasma bPL than controls (10 and 60 min after birth). Our results indicated that differential patterns of secretion of bPL and bPSPB into the maternal and foetal systems occurred at distinct stages of gestation, and these were associated with altered conceptus development after in vitro embryo manipulations, indirectly demonstrating deviations in placental function in IVP pregnancies.

Epithelioid trophoblastic tumor: clinicopathological features with an emphasis on uterine cervical involvement.

We report on the clinical and histological features of five cases of epithelioid trophoblastic tumor, with an emphasis on its involvement of the uterine cervix. All five patients were of reproductive age (median age 38.4 years) and all, except one, presented with vaginal bleeding 3 to 18 years after the most recent pregnancy. One patient presented with amenorrhea. Elevation of serum human chorionic gonadotropin (hCG) was seen in four cases. Pathologically, the tumor involved endocervix in three cases and involved uterine corpus in another two. All five tumors were invasive, nodular lesions consisting of epithelioid intermediate trophoblastic cells that were mononuclear with abundant eosinophilic cytoplasm, along with zones of hyaline material and necrotic debris. In three cases of cervical involvement, the neoplastic cells focally replaced endocervical surface and glandular epithelium, simulating high-grade squamous intraepithelial lesions. Immunohistochemically, all five tumors displayed focal positivity for human placental lactogen and hCG. Positive nuclear staining of p63 was seen in all five cases. All patients received total hysterectomy and various regimes of adjuvant chemotherapy. Three patients survived the tumor with no recurrences or metastases with follow-up periods of 3, 7 and 16 years. One patient is currently alive with lung metastasis 1 month after the surgery. One patient died of tumor metastasis 8 months after the diagnosis. In summary, with its unusual ability to simulate an invasive squamous cell carcinoma and other epithelioid neoplasms, epithelioid trophoblastic tumor frequently poses a diagnostic challenge, especially when involving the uterine cervix. High index of suspicion and an awareness of elevation of serum chorionic gonadotropin are crucial in reaching a correct diagnosis.

Tumor trofoblástico del lecho placentario / Placental site trophoblastic tumor

El tumor trofoblástico del lecho placentario es un tumor excepcional, con no más de 100 casos referidos en la bibliografía. Se trata de una enfermedad neoplásica del trofoblasto intermedio, con capacidad metastatizante en pocos casos. Se presenta el caso de una paciente de 30 años, con una amenorrea de 10+4 semanas y que acudió a urgencias maternales aquejada de sangrado vaginal. Las exploraciones, bioquímica y ecografía fueron compatibles con el diagnóstico de enfermedad trofoblástica, y después de varios legrados ya apareció este raro diagnóstico anatomopatológico. Tras recidiva tumoral se decide histerectomía simple previo estudio de extensión tumoral negativo, y el diagnóstico de tumor trofoblástico del lecho placentario se confirmó en la pieza. Un año después la paciente sigue clínica y analíticamente libre de enfermedad

Placental-site trophoblastic tumor is exceptionally uncommon, with no more than a hundred cases reported in the literature. It is a neoplastic disorder of the middle trophoblast, with metastatic potential in a few cases. Our patient was a 30-year-old woman with amenorrhea of 10 weeks and 4 days' duration who attended the maternity emergency service complaining of vaginal bleeding. Physical examination, laboratory tests and ultrasonographic findings were compatible with a diagnosis of trophoblastic disease. After several curettages, this rare histopathological diagnosis was made. The tumor recurred and negative tumoral extension study and simple hysterectomy were performed. Analysis of the surgical specimen confirmed the diagnosis of placental-site trophoblastic tumor. After 1 year, laboratory tests show no recurrence and the patient is clinically disease-free

Placental villotrophoblastic pulmonary emboli after elective abortion: immunohistochemical diagnosis and comparison with ten control cases.

Although pulmonary trophoblastic embolism is now considered a physiologic phenomenon of normal pregnancy, this phenomenon has not been demonstrated in a living asymptomatic patient. Recently we encountered a 26-year-old woman suspected of pulmonary embolism of villotrophoblastic tissues after therapeutic abortion. Although her serum beta-hCG was low, a computed tomography scan showed multiple nodules in both lungs. Histological examination of a nodule in a lung-biopsy specimen showed granulation tissue surrounding a hemorrhagic mass within which were structures resembling degenerating chorionic villi. Immunohistochemical study on the patient's lung nodule, and a second endometrial-curettage specimen, six control endometrial and tubal specimens containing degenerating chorionic villi, and four endometrial specimens containing viable chorionic villi were performed. The patterns of immunostaining for cytokeratin, human chorionic gonadotropin, human placental lactogen, placental alkaline phosphatase, and inhibin-alpha of the chorionic villus-like structures in the lung nodule were almost identical to those in the degenerating chorionic villi, but different from those of viable villi. This is a unique case of embolism of chorionic villi and trophoblast to the lung in a living patient after therapeutic abortion.

Breast carcinomas with choriocarcinomatous features: case reports and review of the literature.

Breast cancer with choriocarcinomatous features is rare. This report describes four cases of breast cancer with choriocarcinomatous features. The tumor cells were positive for human placental lactogen (hPL) and human chorionic gonadotropin (hCG) by immunohistochemistry. The cases reported in the literature had a poor prognosis and the patients died within a few months after the diagnosis. In this series, two cases were lost to follow-up, but the other two have had disease-free survival for 2 and 4 years, respectively.

Epithelioid trophoblastic tumor of the uterus: cytological and immunohistochemical observation of a case.

Epithelioid trophoblastic tumor (ETT) is a new entity of trophoblastic tumor and 14 such cases were reported by Shih and Kurman in 1998. However, only three subsequent cases supporting ETT have been reported. Recently, we experienced a case of ETT in a 37-year-old woman whose preoperative endometrial brushings showed atypical mononucleate giant cells and who underwent hysterectomy with the diagnosis of a uterine fibroid. The specimens revealed a 2.5 x 3.0 cm yellow-tan intramural nodule located in the lower uterine segment, which was composed of a neoplastic proliferation of intermediate trophoblasts in epithelioid arrangements. Immunohistochemically, the tumor cells were diffusely positive for cytokeratin and inhibin-alpha, and focally positive for human chorionic gonadotropin and human placental lactogen. She presented an uneventful clinical course as of September 2001.

Isolation and characterization of a bovine blastocyst-derived trophoblastic cell line, BT-1: development of a culture system in the absence of feeder cell.

We established a trophoblastic cell line, bovine trophoblast-1 (BT-1), derived from in vitro matured and fertilized blastocyst. While several trophoblastic cell lines have been previously reported using feeder cell, BT-1 could be cultured in the absence of feeder cell. BT-1 was cultured for more than 18 months (over 75 passage) in the absence of feeder cells, using bovine endometrial fibroblast-conditioned medium (fibroblast-conditioned medium). We found that the cell growth was accelerated in fibroblast-conditioned medium. In bromodeoxyuridine incorporation analysis, BT-1 cells growth rate in fibroblast-conditioned medium was about two-fold higher than that in conventional medium. Furthermore, fibroblast-conditioned medium accelerated attachment of BT-1 cells to culture dishes following plating. BT-1 showed epithelial morphology and expressed cytokeratin. During continuous culture, cells accumulated fluid under the cell sheet and form dome-like structure that eventually transformed into free floating vesicles. Reverse transcription polymerase chain reaction analysis and immunoblot analysis demonstrated that BT-1 cells expressed interferon-tau as well as placental lactogen (PL). Immunofluorescence analysis demonstrated that a small number of cells were PL-positive, and these cells were binucleate. The BT-1 trophoblastic cell line could serve as a powerful model system for the study of trophoblast cell lineage and proliferation.

Expression of placental lactogen and cytokeratin in bovine placental binucleate cells in culture.

Binucleate cells are present in ruminant placenta and play an endocrine role in the production of many hormones during pregnancy. We isolated and cultured binucleate cells from bovine placenta at middle to late gestation and characterized these cells using immunofluorescence techniques. Enriched preparations of binucleate cells were obtained using Percoll density gradient centrifugation following collagenase digestion. Binucleate cells in culture preferentially attached to collagen-coated dishes rather than to noncoated plastic dishes. The cells gradually extended their edges on collagen substrata, and finally assumed a flattened morphology. Antibodies to placental lactogen (PL) and pregnancy-associated glycoprotein-1 (PAG-1) specifically stained the majority of round binucleate cells, but not the flat cells. We found that PL-positive binucleate cells were consistently devoid of cytokeratin. In contrast, cytokeratin was expressed in PL-negative binucleate cells as well as mononuclear epithelial cells. Furthermore, the PL-negative flat binucleate cells also developed intense cytokeratin networks in the cytoplasm. These results indicate that cytokeratin expression is inversely proportionate to that of PL in cultured binucleate cells. We conclude that downregulation of cytokeratin in binucleate cells is a function of the state of cellular differentiation.