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PubMed1987

[A simple rapid method of culturing chorionic villi. Identification and description of maternal cells in culture].

Chorionic villi cell cultures is a complement to direct chromosome analysis. It is indispensable for the determination of certain enzymatic activities. A rapid, simple and reliable method of culture is described which allows height quality karyotyping in a week. Confusion with maternal cells is a possible source of error. The authors identified and described these maternal cell types.

Cells, Cultured↗
PubMed2004

Isolation and characterization of mesenchymal progenitor cells from chorionic villi of human placenta.

BACKGROUND: BM-derived mesenchymal stem cells (MSC) are attractive sources for autotransplantation with no risk of rejection, but the use of these cells bas problems, including the necessity of harvesting BM from donors, the donors' age-dependency, limitation to autologous use and difficulty of use for patients with hereditary diseases. We report a method of isolating placenta-derived mesenchymal progenitor cells (PDMPC) that can be used as an alternative source of MSC. METHODS: We isolated PDMPC from human fetal chorionic villi using the explant culture method, from placentas collected after neonatal delivery (38-40 weeks of gestation). The PDMPC were characterized by morphologic and immunophenotypic analysis. The differentiation ability of mesenchymal and neural lineages was detected using specific culture conditions and determined by morphology, reverse transcription(RT)-PCR, histochemical staining and immunocytostaining. RESULTS: The PDMPC all originated from fetal chorionic villi, as confirmed by fluorescence in situ hybridization analysis. The PDMPC population consisted of spindle-shaped cells and large flat cells. The PDMPCexpressed CD13, CD44, CD73, CD90, CDIO5 and HLA class I as surface epitopes, but not CD31, CD34, CD45 and HLA-DR. These cells differentiated into osteocytes, chondrocytes and adipocytes under specific culture conditions, and were also induced to form neural-like cells. DISCUSSION: Our study shows that PDMPC can differentiate into mesenchymal lineages and be induced to form neural-like cells. Thus, PDMPCisolated from chorionic villi of placenta may provide a novel source for the research of stem and progenitor cells in placenta, cell therapy and regenerative medicine, particularly as a source of allogenic mesenchymal stem and progenitor cells with little ethical conflict and various advantages

Antigens, CD↗
PubMed1986

Cytogenetic analysis of chorionic villi: a technical assessment.

Eighty-five samples of chorionic villi from women undergoing prenatal diagnosis at 8 to 12 weeks' gestation were subjected to cytogenetic analysis. Samples were prepared by a direct technique that permits limited analysis within two hours and by a short-term culture technique that permits detailed structural analysis within one week. An adequate number of cell divisions for cytogenetic analysis was obtained from 96% of living fetuses. Using both the direct technique and short-term culture, satisfactory banded chromosomal preparations were made in 93% of cases. Eleven of 12 pregnancies (92%) shown by ultrasound to be dead shortly before sampling, had cytogenetic abnormalities. Further studies are needed to develop banding definition equivalent to that available on cultured amniocytes.

Adult↗
PubMed1989

[Chorionic villi sampling. Results with the first 500 samples].

531 chorionic villi forceps biopsies were carried out, to look for genetic diseases. Sampling was impossible in 24 cases (4.5%). The following indications were chosen: maternal age over 38 years: 361 cases; parental chromosome abnormality: 28 cases; previous history of chromosome abnormality: 21 cases; X-linked disease: 77 cases; metabolic disease: 32 cases; hemoglobinopathy: 7 cases; blood group determination: 5 cases. An elective abortion was carried out in 44 cases (8.7%), whereas 465 pregnancies were not interrupted. Full term has been reached in 327 pregnancies. The rate of miscarriage in this group is 3.9% (13 miscarriages out of 327 pregnancies followed). The results obtained at the Saint-Vincent-de-Paul Hospital show that the cytogenetic technique is reliable, and that the low risk of sampling allows its application in the diagnosis of chromosome anomalies.

Adult↗
PubMed1985

[Studies on mode of cystine aminopeptidase production in the human placental chorionic villi and its effect on angiotensin II during pregnancy].

The production mechanism and physiological significance of cystine aminopeptidase in human placental chorionic villi were investigated with the following findings: The CAP activity of the lysosome fraction was the greatest (p less than 0.05) in each trimester. The CAP activity determined using Kato's method, which can separate lysosome better, was significantly (p less than 0.05) lower than in our lysosome fraction. CAP activity increased significantly after 30 min. (p less than 0.05) in a human chorionic villi culture continued to rise up to 60 min. (p less than 0.01) and then fell slightly. In the presence of angiotensin II, CAP activity increased rapidly for the first 30 min. and after 60 min. fell rapidly and continuously (p less than 0.01). With ADP and PGF2 alpha added it decreased insignificantly. With galactose and sialic acid, the increase was significant only at 30 min. after adding galactose. Thin layer chromatography of angiotensin II after incubation with partially purified CAP for 3 hr at 37 degrees C indicates that CAP splits angiotensin II. A chromatogram of the amino acid shows that 76n mol aspartic acid, 30n mol valine, 49n mol isoleucine, 22n mol tyrosine, 55n mol phenylalanine and 54n mol arginine separated from 1 mumol angiotensin II. These results suggest that CAP originates in other lysosomeseized granules and is an angiotensinase-like enzyme. Since it acts like angiotensinase, it is probably one of the factors maintaining the mother's blood pressure at normal levels during pregnancy.

Amino Acids↗
PubMed1992

[Intraplacental hematoma following chorionic villi biopsy].

One of the most typical complications following chorionic villae sampling (CVS) is vaginal bleeding shortly or some days after the intervention, resulting in abortion in some cases. We report on a case of intraplacental haematoma (pathology: massive subtotal acute haemorrhagic placental infarct), followed by acute placental insufficiency in the 29th week of pregnancy. Therefore, we decided on an emergency caesarean section.

Adult↗
PubMed1990

Fetomaternal transfusion depends on amount of chorionic villi aspirated but not on method of chorionic villus sampling.

Transcervical and transabdominal chorionic villus sampling are believed, on the basis of indirect evidence, to result in fetomaternal transfusion. We sought to measure this phenomenon by devising a simple method that would allow us to identify variables that influence fetomaternal transfusion. We investigated patients undergoing transcervical-chorionic villus sampling (n = 15) and transabdominal-chorionic villus sampling (n = 15), restricting the sample to subjects who required only a single catheter passage or needle insertion to obtain villi. Maternal serum alpha-fetoprotein was measured before and after the procedure along with alpha-fetoprotein concentration of the transport medium into which the villi had been aspirated. We first confirmed that the change in maternal serum alpha-fetoprotein levels after chorionic villus sampling, an indirect measure of fetomaternal transfusion, was indeed correlated with the alpha-fetoprotein concentration of transport medium into which the villi were aspirated (p = 0.0350). Fetomaternal transfusion next proved to be correlated with the amount of villi obtained (p = 0.0279). However, when adjusted for the amount of villi obtained, no significant difference was observed between transcervical and transabdominal-chorionic villus sampling with respect to the change in maternal serum alpha-fetoprotein levels after chorionic villus sampling (p = 0.8512). These data suggest that the magnitude of fetomaternal transfusion depends on the amount of villi obtained but not on the chorionic villus sampling method used.

Adult↗
PubMedDate not supplied

Mechanisms of regulation of the capillary bed in the human chorionic villi.

The purpose of the present researches was to confirm the hypothesis that in the human placenta the microcirculation is subjected to humoral control, because of the absence of innervation in the chorionic villi and the ability of the capillaries to contract themselves. Chorionic villi were incubated in presence of vasoactive substances, Histamine, Serotonin, Bradykinin, Dopamine, Enkephalin Prostaglandins. After incubation, the specimens were fixed and embedded in resin. Semithin sections were submitted to a computerized analysis for the evaluation of the ratio between surface of capillaries and surface of the villus. The results confirmed that in the control of capillaries in the human placenta most of the tested substances show a specific action, dose- or time-dependent.

Capillaries↗
PubMed1994

Chromosome preparations of both direct and long-term cultures from the same fragment of chorionic villi.

A simplified method is described for processing both direct preparations and long-term cultures from the same fragment of chorionic villi. Enzyme separation of the outer trophoblast layers (used for direct preparations) from the inner mesenchymal core (used to initiate long-term cultures) facilitates the utilization of the same fragments for the two procedures, without jeopardizing the success of either method. This has proved useful in cases where the sample was so small that only one method of chromosome preparation may have been possible using other techniques.

Cells, Cultured↗
PubMedDate not supplied

Short communication: trophoblast proliferation is increased in chorionic villi from pregnancies with fetal trisomy 18.

Pregnancies with fetal trisomy 18 suffer from severe intrauterine growth restriction from the first trimester. This cross-sectional study of chorionic villi obtained from ongoing singleton pregnancies at 10-14 weeks of gestation, semiquantitatively examines chorionic villus stromal and trophoblastic cell proliferation rates in ongoing chromosomally normal and trisomy 18 pregnancies. Chorionic villi were stained using a standard immunohistochemistry protocol with a monoclonal antibody to the Ki-67 antigen and the number of Ki-67 positive trophoblastic and stromal cells per villous cross-section was compared between groups. The number of Ki-67 positive trophoblastic and stromal cells was significantly higher in the group (n=10) with trisomy 18 compared to the chromosomally normal group (n=25;t=2.1, P=0.03 and t=5.3, P<0.001 respectively). Abnormalities in control of cell turnover during development may be important in the pathogenesis of the clinical features of trisomic pregnancies.

Adult↗
PubMedDate not supplied

Comparative study of 15 lysosomal enzymes in chorionic villi and cultured amniotic fluid cells. Early prenatal diagnosis in seven pregnancies at risk for lysosomal storage diseases.

A large number of chorionic villi samples obtained from women undergoing elective first trimester termination of pregnancy was analysed by enzyme assays similar to those applied to cultured amniotic cells. The levels of 15 lysosomal enzymes were compared to those observed in tissue cultures of amniotic cells obtained through amniocentesis at 16-18 weeks of pregnancy and the results were discussed in order to assess the usefulness of trophoblast biopsy for first trimester diagnosis of hereditary lysosomal diseases. The data suggest the applicability of this source of fetal cells for prenatal diagnosis of fifteen respective genetically determined enzyme deficiencies with the probable exception of alpha-L-iduronidase deficiency. Enzyme determinations were performed on chorionic villi samples of two pregnancies at risk for Tay-Sachs disease, three pregnancies for GM1 gangliosidosis type 1, one for mucopolysaccharidosis type VI and one for Wolman's disease.

Amniocentesis↗
PubMed1993

The value of histomorphological features of chorionic villi in early spontaneous abortion for the prediction of karyotype.

This study was conducted to evaluate the value of histomorphological features of chorionic villi, such as size, shape, vascularity, trophoblast proliferation and trophoblastic pseudo-inclusions, for the prediction of chromosomal abnormality. Slides of 83 early spontaneous abortions were assessed by three observers. Assessments and karyotype were compared and likelihood ratios computed. Likelihood ratios of < or = 0.40 or > or = 2.50 were not obtained by all three observers for any of the features. One or two observers obtained likelihood ratios of > or = 2.50 for lacunar stromal hydrops, presence of trophoblastic hyperplasia, moderate to abundant trophoblastic hyperplasia, presence of trophoblastic lacunae, few intervillous fibrin deposits and few intervillous trophoblastic buds. Likelihood ratios of < or = 0.40 were found for small chorionic villi and presence of basophilic staining. Lacunar stromal hydrops and trophoblastic lacunae were predictive of triploid karyotype, but not specific for any other type of chromosomal abnormality. After application of data previously obtained on Cohen kappas, lacunar stromal hydrops, moderate to abundant trophoblastic hyperplasia and presence of trophoblastic lacunae remained as possibly useful features, again mainly for identifying triploidy. Most of the items claimed to be related to abnormal karyotype, however, were not predictive at all.

Abortion, Spontaneous↗
PubMed1985

Incomplete X chromosome dosage compensation in chorionic villi of human placenta.

Studies of glucose-6-phosphate dehydrogenase (G6PD) in heterozygous cells from chorionic villi of five fetal and one newborn placenta show that the locus on the allocyclic X is expressed in many cells of this trophectoderm derivative. Heterodimers were present in clonal populations of cells with normal diploid karyotype and a late replicating X chromosome. The expression of the two X chromosomes was unequal, based on ratios of homodimers and heterodimers in clones. Studies of DNA, digested with Hpa II and probed with cloned genomic G6PD sequences, indicate that expression of the locus in chorionic villi is associated with hypomethylation of 3' CpG clusters. These findings suggest that dosage compensation, at least at the G6PD locus, has not been well established or maintained (or both) in placental tissue. Furthermore, the active X chromosome in these human cells of trophoblastic origin can be either the paternal or maternal one; therefore, paternal X inactivation in extraembryonic lineages is not an essential feature of mammalian X dosage compensation.

Cells, Cultured↗
PubMed1996

Proliferation kinetics of chorionic villi in chromosomally normal and abnormal spontaneous abortions analyzed by premature chromosome condensation and northern blot.

The activity of cell proliferation in human chorionic villi (CV) obtained from early spontaneous abortions (SAB) and from elective abortions (EAB) was determined by means of premature chromosome condensation (PCC), and by Northern blot analysis of expression of proliferation antigens Ki67 and PCNA. The rate of chromosome aberrations among the SABs was 57%, while the EABs, taken as controls, were shown to be chromosomally normal. PCCs were induced by fusion of the chorionic interphase cells with mitotic Chinese hamster ovary (CHO) cells. To analyze the proliferation stages of the chorionic G1-interphases, the potential proliferation index (PPI) was ascertained. The PPI values found in SABs ranged from 36% to 97% as described for intensely proliferating tissues. They did not differ from the values found in the controls (34-90%), indicating maintenance of proliferation activity of CV cells even after death of the embryo in missed abortions. No significant PPI differences were observed between the abortion groups with different cytogenetic results. The expression of proliferation associated antigens Ki67 and PCNA showed no significant differences between mean values of the total of SABs and of the controls. However, the comparison of mean values of chromosomally abnormal with chromosomally normal SABs revealed a significantly reduced Ki67 activity in the SABs with chromosome aberrations. A similar results was obtained when comparing mean values of Ki67 and PCNA expression from trisomic SABs with those of the controls. By further subdivision of chromosome aberrations a decrease for the expression of both antigens in chorionic villi from early lethal trisomies, and a significantly increased Ki67- and PCNA-expression in triploidies became evident.

Abortion, Induced↗
PubMed1990

A confocal and conventional epifluorescence microscope study of the intermediate filaments in chorionic villi.

An immuno-epifluorescence microscopic study of first trimester human chorionic villi has revealed different patterns of distribution for intermediate filament proteins. Keratin staining was restricted to the trophoblastic epithelium. The protein was shown to be concentrated into desmosome-containing apical and basal cytoplasm. Strands of keratin-rich cytoplasm extended across the interior of the syncytium, integrating basal and apical layers. This arrangement integrates the epithelial cytoskeleton in a manner well adapted to resisting shearing forces. Vimentin was found in the cells of the villous core and was most strongly expressed in endothelial cells. Desmin was also restricted in distribution of spindle-shaped cells of the villous core. It was excluded from terminal villi.

Chorionic Villi↗
PubMed1989

[In situ hybridization of chorionic villi cells].

Two cloned alpha fragments of satellite repetitive DNA sequences were used to modify the method of DNA hybridization in situ (HIS) for tissue cells of human chorionic villi. Techniques for the cells of short-term lymphocytic cultures were used to optimize the conditions of HIS in chorionic villi cells with regard to a more specific localization of cloned repetitive DNA sequences in the target homologous chromosomes. A total of 262 specimens were prepared using the HIS method. As the results show, both cloned fragments are well applicable for routine cytogenic practice, the fragment p YAM 10-40 for the number of chromosomes, and fragment p YAM 9-72 for chromosomes 18.

Cells, Cultured↗
PubMed1980

Ultrastructure of the epithelium of the chorionic villi of the human placenta.

Human placenta samples obtained at 4, 6, 8 and 10 weeks of gestation and at full term were studied by transmission electron microscopy (TEM). The chorionic villi at early stages of gestation exhibited abundant microvilli on the free surface of the syncytium. The chorionic villi at full term were covered by syncytium which bore short microvilli on its free surface. The syncytial cells were remarkably higher in density than Langhans' cells. In the earlier period of gestation, a dividing membrane was noticed in the inside of the syncytial cells. The cytoplasm of the syncytial cell contained comparatively small mitochondria, well-developed endoplasmic reticulum cells, ill-developed Golgi systems, electron-dense lipoid droplets and plenty of ribosomes. A great number of Langhans' cells were noticed in early gestation; only very few were noticed the later part. Mitochondria of Langhans' cells were comparatively large and the Golgi system was well-developed in the first trimester. Endoplasmic reticulum was less well developed and there were few ribosomes. The basement tended to become thicker a pregnancy proceeded and lamellar structures appeared on its inner face.

Chorionic Villi↗
PubMed1987

Direct chromosome preparations from chorionic villi: a method for obtaining extended chromosomes and recognizing mosaicism confined to the placenta.

A modified technique is described for obtaining extended metaphase chromosomes (600 bands) from direct preparations of chorionic villi, allowing rapid detection of subtle structural aberrations with minimal risk of maternal-cell contamination. Analysis of multiple individually processed villus fragments may be helpful in recognizing mosaicism confined to chorionic villi.

Chorionic Villi↗