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PubMed1994

Cytogenetic analysis of chorionic villi for prenatal diagnosis: an ACC collaborative study of U.K. data. Association of Clinical Cytogeneticists Working Party on Chorionic Villi in Prenatal Diagnosis.

A prospective 3-year collaborative study was undertaken in 1987 to collect cytogenetic data from diagnostic chorionic villus samples (CVS) in the U.K. in order to determine the predictive value of the chromosome abnormalities encountered. Twenty-seven laboratories contributed a total number of 7595 cases, of which 97.6 per cent were successful. Excluding single cell anomalies, a total of 480 cytogenetic abnormalities were reported, of which 137 were familial structural rearrangements and 343 were de novo problems. Non-mosaic trisomies of chromosomes 13, 18, and 21 (n = 157), non-mosaic sex chromosome abnormalities (n = 33), and triploidy (n = 6) were all confirmed in cells of fetal origin where follow-up information was available. Of the nine remaining non-mosaics including tetraploidy, trisomies of other autosomes, and extra markers, only a trisomy 16 and a case of a supernumerary marker proved genuine. Eighty-eight cases of mosaicism were reported to the study, of which only nine were confirmed as genuine: two cases involving chromosome 13, one trisomy 18, two examples of extra marker chromosomes, three 45,X, and one 47,XXX. There were no reports of false-negative findings. Presumptive maternal cell contamination was encountered in 39 cases, a detected incidence of 0.5 per cent. Four cases of presumptive 'vanishing twin' were recorded: in three of these, direct preparations showed a female karyotype, whereas culture indicated a male (with male fetuses in two cases). The fourth case was of a female fetus with male and female cells in the CVS cultures. Subtle structural chromosome abnormalities were missed in three instances. Accurate prediction of the fetal karyotype was shown to require detailed knowledge of both the nature and the distribution of abnormal cells in the extra-embryonic tissues. In many cases, this could only be made where results from direct preparations and cultured cells were available. A number of conclusions were reached from these and similar data in the literature regarding the reliability of chromosome findings in CVS.

Chorionic Villi Sampling↗
PubMed2001

The diagnostic performance of cytogenetic investigation in amniotic fluid cells and chorionic villi.

First-trimester chorionic villus sampling has not reached the popularity of second-trimester amniocentesis in prenatal cytogenetic diagnosis, in contrast to initial expectations. We investigated whether a difference in the diagnostic performances of cytogenetic investigation in amniotic fluid (AF) cells and chorionic villi in favour of AF-cells might justify this. Diagnostic performance was measured as laboratory failure rate, karyotype quality (G-band score, rate of follow-up samples, rate of wrong diagnoses), and karyotype representativity (rate of follow-up samples, rate of wrong diagnoses). From 1993-1999, 11 883 AF-samples were investigated (AF-cells). In chorionic villi, short term culture preparations solely were karyotyped from 1993-1996 (n=3499) (STC-villi), short and long-term culture preparations simultaneously provided a sufficient amount of tissue being available from 1997 onwards (n=1829) ((STC+LTC)-villi). Laboratory failure rates were the same after amniocentesis (0.40%) and chorionic villus sampling (0.50%). G-band scores (mean+/-SD) were equal in AF-cells (373+/-38.1) and LTC-villi (364+/-32.6) but significantly lower in STC-villi (311+/-34.6) (p=0.001). Follow-up sampling rates because of quality reasons were the same in AF-cells (0.14%), STC- villi (0.13%) and (STC+LTC)-villi (0.11%). Two wrong diagnoses turned up among AF-cells. Follow-up sampling rates because of representativity reasons differed significantly between AF-cells (0.10%), (STC+LTC)-villi (1.31%), and STC-villi (1.99%) (p<0.001). However, the ratios of the total numbers of follow-up samples and uncertain or abnormal cytogenetic results in STC, and (STC+LTC)-villi at cytogenetic risks > or =3% (0.132 and 0.160, respectively) were equal to that in AF-cells at risks <3% (0.155). Two wrong diagnoses were made in STC-villi. Diagnostic performance improved in the rank order of STC-villi, (STC+LTC)-villi and AF-cells. At cytogenetic risks > or =3%, (STC+LTC)-villi showed a diagnostic performance equal to that in AF-cells. This might justify a selective use of chorionic villus sampling.

Amniocentesis↗
PubMed2000

Four years' cytogenetic experience with the culture of chorionic villi.

In 1958 chorionic villus samples, investigated by culture method, we found 137 (7%) abnormalities. The abnormal results were classified in certain abnormal (generalised abnormal at high probability) and uncertain abnormal (potentially confined to the placenta) results. Certain abnormal were 73 cases (3.7%). Uncertain abnormal were 64 cases (3.3%), in which confirmation studies were done in 47 cases. In 12 cases of these 47, the abnormality was confirmed and in 35 cases (1.8%) the abnormality was confined to the placenta. Among the latter cases, poor pregnancy outcome [16% intrauterine death (IUD), 6% intrauterine growth retardation (IUGR)] was increased. Total maternal cell contamination was not seen. The positive predictive value of all confirmed abnormal cases was 66%. The positive predictive value was 100% for indications 'ultrasound abnormalities' and 'carrier' and between 50 and 60% for all other indications. Predictive value among uncertain abnormal cases was low (26%). However, the positive predictive value depends of the type of abnormality. Therefore we conclude that the culture method for chorionic villi is a good test for indications 'ultrasound abnormalities' and 'carrier' and reliable for all other indications. Whether or not follow-up investigations should be offered to the parents depends of the type of abnormality. We conclude that the culture method is reliable for prenatal diagnosis and can be used as the sole investigative method.

Chorionic Villi Sampling↗
PubMed1987

[Biopsy of the chorionic villi].

The biopsy of chorionic villi (BCV) has by now been applied in approximately 20,000 pregnant women world-wide in prenatal diagnosis. The first experience in the Federal Republic of Germany was gained in the centres supported by the Federal Ministry for Research and Technology. Based on a questionnaire sent to each of these centres, a survey of the situation is given up to July 1986; at that time, each centre had an average of 2 years of experience with this new diagnostic method. At the present time, approximately 10% of all amniocenteses are replaced by BCV, the transcervical, ultrasonic catheter control method being applied in most cases. In 89%, BCV also resulted in a reliable genetic diagnosis. Bleedings without induction of abortion (16.7%) were relatively frequent complications. Prediagnostic abortions occurred in 10.6% and 5.5% following BCV. However, every 5th abortion took a febrile course. Genetic diagnoses confirmed an evidently normal finding in 87.9% and a pathological finding in 4.4%. 6% of the findings made a control necessary. Abortion was performed in 3.8% of the cases following biopsy of chorionic villi.

Abortion, Eugenic↗
PubMed1993

Immunophenotyping of mitotic cells from long-term cultures of chorionic villi.

Chromosomal mosaicism in chorionic villus samples (CVS) may arise from different sources, such as clonal diversity within the chorionic tissue or contamination with maternal cells. To determine the origin of karyotyped cells, we compared the immunocytochemical features of mitotic cells in CVS long-term cultures with histological sections of their tissue of origin, i.e. chorionic villi. Immunolabelling of intermediate filaments specific for epithelial cells (cytokeratin) and mesenchymal cells (vimentin) established that mitoses yielded from CVS long-term cultures indeed stem from independently growing clones derived from both the epithelial and mesenchymal parts of the chorionic villi. Thus, mosaicism in CVS cultures may reflect true genetic heterogeneity within the biopsy. However, epithelial chorionic cells undergo in vitro metaplasia leading to co-expression of cytokeratins and vimentin. Fetal-specific immune markers (beta-HCG and SP1-glycoprotein) are not reliably expressed in CVS cell culture.

Chorionic Gonadotropin↗
PubMed2005

Prenatal diagnosis of cystinosis by quantitative measurement of cystine in chorionic villi and cultured cells.

OBJECTIVE: Prenatal diagnosis of cystinosis has been available for over 30 years by the incubation of cultured amniotic cells, intact chorionic villi and cultured chorionic cells with [35S]-cystine followed by thin layer chromatography and visual inspection of autoradiographs of the chromatograms for cystine. This method has proved highly reliable but because of the short half-life of [35S]-cystine, its cost and the length of the assay procedure, an alternative method of diagnosis was investigated. METHOD: Cystine was quantitatively measured in chorionic villi directly, in cultured chorionic villi and cultured amniotic cells using a cystine-binding protein from Escherichia coli. RESULTS: Twelve pregnancies at risk for cystinosis were monitored by both the [35S]-cystine uptake method and the new quantitative method in uncultured chorionic villi. There was no discrepancy between the results obtained with the two methods and subsequently 15 pregnancies have been monitored by the quantitative assay only--13 in chorionic villi directly, 1 in cultured chorionic villi cells and 1 in cultured amniotic cells. Grossly elevated levels of cystine were found in seven pregnancies. CONCLUSION: An unequivocal diagnosis of cystinosis can be made within 24 h of sampling by the quantitative measurement of cystine in uncultured chorionic villi.

Abortion, Eugenic↗
PubMed1991

[Research on karyotypic consistency between chorionic villi and fetus].

In the last decade, chorionic villi sampling (CVS) has been widely accepted as a new technique in prenatal diagnosis. An inconsistency in karyotypes between CVS and fetal tissue was found in clinical practice, although the chorionic villi and the fetal tissues were from the same fertilized ovum. Karyotypes from direct chromosomal preparation of chorionic villi and cultured fetal tissues in 52 artificial abortion samples were compared. Results showed two cases with karyotypic inconsistency: (1) 46, XX/46, XY in the chorionic villus direct preparation and 46, XX in the fetal tissue; (2) 46, XY in the direct preparation of chorionic villi and 45,XY, t (15q21q) de novo in the fetal tissue. Our research demonstrates that karyotype from chorionic villus direct preparation does not always reflect the fetal karyotype.

Adult↗
PubMed1988

Chromosome analysis in chorionic villi samples from the first trimester elective terminations.

Chorionic villi sampling (CVS) has become a first trimester alternative to amniocentesis for prenatal diagnosis. The cytogenetic findings in 150 experimental samples are presented. The ages of the mothers ranged from 12 to 35 years, but the majority of them were 18 and 19 years of age. Various parameters of culturing and processing the samples in order to improve the method, were investigated. Short term incubation for 48 h was the method routinely employed in processing the biopsies for cytogenetic analysis. In the first series of 100 cases one mosaic case (46,XX/45,X), one Robertsonian translocation (13;14), one marker chromosome and one fragment were found. The foetal tissues were not analysed for chromosomes. In the second series of 50 samples, one case of mosaicism was found in the chorionic villi (46,XX/47,XX, 18q-), but this abnormality was absent in the foetal tissue. One variant inv(9) was observed in the foetal tissue as well as in the chorionic villi. In all other cases the karyotypes from the chorionic villi samples matched those of the corresponding foetal samples. There was no maternal contamination in this series of 50 samples. The discrepancies in the cytogenetic results from other investigators are discussed.

Chorionic Villi↗
PubMed1983

Absence of maternal contamination of chorionic villi used for fetal-gene analysis.

Chorionic villi can be obtained by direct transcervical aspiration at 9 to 10 weeks' gestation and used for analysis of fetal DNA. However, for the method to be reliable, there must be no detectable contamination by maternal DNA. To investigate the question of contamination, we compared the DNA of chorionic villi from five fetuses with that obtained from maternal lymphocytes, using the restriction endonuclease Taql and specific DNA probes for a pair of alleles on the X chromosome. The alleles yield fragments of different lengths when digested with Taql (length polymorphism), which can be demonstrated by electrophoresis and hybridization with the radioactive DNA probes. If the pattern obtained with the chorionic DNA is different from that obtained with the maternal DNA, contamination is not present. In two cases the fetal DNA of the chorionic villi was shown to be uncontaminated by maternal tissue. In one of these cases the mother was heterozygous and the fetus was homozygous; in the other the mother was homozygous and the fetus was heterozygous. In three other cases no definitive conclusions could be drawn, because the genotypes of the fetus and mother were identical. We conclude that chorionic villi at 9 to 10 weeks' gestation are a source of fetal DNA that can be used for gene analysis, with no detectable contamination by maternal DNA.

Alleles↗
PubMed1987

Maternal cell contamination in cultured chorionic villi: comparison of chromosome Q-polymorphisms derived from villi, fetal skin, and maternal lymphocytes.

Maternal cell contamination of chorionic villi (CV) samples used for first trimester prenatal diagnosis can cause obvious and/or unrecognized diagnostic dilemmas. The purpose of this investigation is to assess the frequency of maternal cell contamination (MCC) in chorionic villus samples and to evaluate selected parameters which might predict where contamination is more likely to have occurred. Maternal lymphocytes, chorionic villi from ultrasonically directed transcervical catheter aspiration, and fetal tissue were obtained at 8-11 weeks gestation from 45 patients undergoing elective termination. Quinacrine (Q) banded metaphases were compared from duplicate direct preparations of chorionic villi; cultured chorionic villi, fetal fibroblast tissue cultures, and maternal lymphocyte cultures. Q-polymorphisms in metaphase chromosomes were 100 per cent concordant between fetal tissue and direct CV preparation. However, evidence for maternal cell contamination occurred in 13.1 per cent of cultured chorionic villi preparations where polymorphisms were found to be identical between maternal and cultured CV and both distinct from fetal tissue preparations. Where MCC was identified, it was noted that CV cell cultivation interval was prolonged (24.2 +/- 6.8 days) compared with non-contaminated cultures (14.1 +/- 4.4 days) (p less than 0.05). We conclude that maternal cell contamination is a significant problem with chorionic villus sampling. Where direct preparations are not employed or when cultures are 'slow growing', MCC may be a significant and unrecognized complication re: fetal diagnosis. Direct preparations, multiple cultures, quinacrine banding, and maternal Q-polymorphism comparisons can minimize diagnostic dilemmas secondary to maternal cell contamination.(ABSTRACT TRUNCATED AT 250 WORDS)

Cells, Cultured↗
PubMed1990

Current method for first and second trimester prenatal diagnosis: transabdominal chorionic villi sampling.

Transabdominal CVS (chorionic villi sampling) was performed in 40 patients belonging to first or second trimester of pregnancy with 97.5% success, using a double and later single needle technique with no immediate complications. Karyotyping by direct method was successful in 75% of samples thus indicating the feasibility of cytogenetic analysis using villus tissue from first and second trimesters of pregnancy. It is a safe, simple, rapid and practical method of prenatal diagnosis in first and second trimesters of pregnancy with a potentially lower risk for fetal and maternal complications.

Chorionic Villi Sampling↗
PubMedDate not supplied

The expression of major histocompatibility antigens in the chorionic villi of molar placentae.

Frozen sections of chorionic villi from molar placentae, ranging in menstrual age from 9 to 19 weeks, were studied by an immunoperoxidase technique using the monoclonal antibody to HLA-A,-B,-C antigens W6/32. The distribution of class I HLA antigens in molar placentae was compared with that in normal placental tissue of comparable menstrual age. The results showed that the distribution of these antigens in molar placentae is similar to that seen in normal placentae. In both tissues, class I HLA antigens were absent from the syncytiotrophoblast and the cytotrophoblast cells of the chorionic villi. However, these antigens were found to be present on cells in the mesenchymal stroma of both normal and molar chorionic villi.

Chorionic Villi↗
PubMed1981

[Studies on influence of synthetic LH-RH and dbcAMP on normal chorionic villi and BeWo cells (author's transl)].

The influence of synthetic LH-RH and dbcAMP on human chorionic villi and BeWo cells was studied through measurement of cAMP, hCG and estradiol. The results obtained were as follow: 1) There was a rapid increase of cAMP in chorionic villi after the 7th week of gestation, marking a maximum value at the 9th week, and gradually decreasing thereafter. 2) When dbcAMP was added to chorionic villi, there was a significant increase estradiol production in chorionic villi and hCG secretion in media. 3) The stimulation of cAMP in chorionic villi and hCG secretion into medial resulting from the addition of synthetic LH-RH to chorionic villi showed significant increases at 5 minutes and 30 minutes, respectively. 4) When dbcAMP or synthetic LH-RH was added to BeWo cells, increased secretion of hCG and estradiol into media was seen. It therefore is concluded that there is a relationship between the activity of synthetic LH-RH in which cAMP participates and a mechanism involving hCG secretion and estradiol production originating from human chorionic villi and BeWo cells. This is turn suggest the possibility of the existence in chorionic villi of a LH-RH like substance which plays a role in the production or secretion of hormone in the placenta.

Bucladesine↗
PubMed1990

Characterization of long-term cell cultures of human chorion villi and fibroblasts using antibodies to cytoskeletal proteins.

In long-term cultures of chorion villi the expression of cytoskeletal proteins--vimentin, cytokeratin, actin, tropomyosin and vinculin--was studied using immunoreactivity to antibodies. Vimentin was detected in all cells of chorionic villi cultures despite their morphological appearance. In comparison to fibroblast cultures from human skin biopsies with bundle-like vimentin structures, chorionic villi cells showed fine ramified vimentin filaments. Some of the chorionic villi cells with typical fibroblastoid appearance also had cytokeratin filaments. No differences were observed in the distribution of vinculin. Tropomyosin structure and actin filaments could be detected in cultures of chorionic villi whereas fibroblasts showed only an intense diffuse staining.

Antibodies, Monoclonal↗
PubMed1990

Variation of lysosomal enzyme activity with gestational age in chorionic villi.

The activities of 14 lysosomal enzymes in chorionic villi at gestational ages of 6-12 weeks were assayed. Arylsulphatases A and B, alpha-glucosidase and beta-glucuronidase activities increased with advancing gestational age. When compared with the activity in cultured amniotic fluid cells, arylsulphatase A, beta-galactosidase, alpha-glucosidase, heparan N-sulphatase, alpha-L-iduronidase, alpha-mannosidase, neuraminidase, and sphingomyelinase showed significant differences. All except beta-glucuronidase showed lower activity in chorionic villi than in cultured amniotic fluid cells. Prenatal diagnosis using chorionic villi was possible except for alpha-L-iduronidase. Storage at -20 degrees C up to 42 days did not significantly affect activity. The results emphasize the importance of using fresh or frozen age-matched control tissue for diagnosis.

Amniotic Fluid↗
PubMed1986

Acceptability of chorionic villi sampling for prenatal diagnosis.

The factors that influence women in choosing between first-trimester chorionic villi sampling and second-trimester amniocentesis for prenatal diagnosis were investigated. Five hundred twenty women of advanced maternal age who had previously undergone prenatal diagnosis by amniocentesis and were delivered of a normal infant were requested to complete a questionnaire concerning their attitudes toward amniocentesis and chorionic villi sampling. The majority of respondents indicated that the time at which chorionic villi sampling is performed (76%), the rapid availability of diagnostic results (72%), and the type of abortion procedure available (68%) would make them choose this method. In contrast, the factors that influenced women to choose amniocentesis included the known low risk of spontaneous abortion (76%) and confidence in the skill of the obstetrician who would perform the procedure (56%). When all factors were considered together, 68% of the respondents chose amniocentesis based on the known low risk of spontaneous abortion, whereas for those who chose chorionic villi sampling (32%), the major criterion was the fact that the procedure is performed in the first trimester. However, 87% of women who preferred amniocentesis indicated that if the risk of spontaneous abortion associated with chorionic villi sampling. These results indicate that for many women of advanced maternal age, the acceptability and the use of chorionic villi sampling will be dependent on the demonstration that the risk of fetal loss is low, approaching that of amniocentesis.

Abortion, Spontaneous↗
PubMed1984

"Map" of proteins resolved from human chorionic villi by two-dimensional electrophoresis.

Two-dimensional electrophoresis was applied to specimens of human chorionic villi obtained during the first trimester of gestation, the object being to simultaneously map several hundred polypeptide gene products. Genetically normal specimens were homogenized in a urea-based denaturant and the supernates were electrophoresed with use of the "ISO-DALT" system. Four categories of proteins are distinguished on the map: previously identified proteins present in chorionic villi and other cell types; unidentified proteins present in chorionic villi and other cell types; proteins present in chorionic villi and amniotic fluid but not in other cell types; and proteins probably originating from the amnio-chorionic plate. The reference map for chorionic villi provided in this study may serve as the basis of determining whether genetic analyses conducted in the first trimester accurately represent the fetal genotype.

Amnion↗
PubMed1989

X-chromosome inactivation in cultured cells from human chorionic villi.

X-chromosome inactivation was investigated in human chorionic villi in the first trimester of pregnancy and cultured cells established from them. Expression of glucose-6-phosphate dehydrogenase (G6PD) was evaluated in these extraembryonic cells from four females heterozygous for the electrophoretic variants (AB) of G6PD. In each case the uncultured villi as well as derived cultured cells expressed the AB phenotype for G6PD with about equal intensity for the A and B bands. Single-cell-derived clones established from two of the four cases expressed either G6PD A or B. One clone expressing G6PD B was fused with mouse cells, and a hybrid clone retaining the inactive human X chromosome was isolated; there was no evidence of human G6PD expression in this clone retaining an inactive human X. DNA methylation in the first intron of the human gene for hypoxanthine phosphoribosyltransferase (HPRT) was evaluated in the four pairs of cultured villi and fetal cells. No differences were detected between the cultured villi and fetal cells as they all showed bands characteristic of an inactive X from somatic cells. These results show that there is no preferential inactivation of an X in the majority of cells that constitute human tertiary chorionic villi or in cultured cells derived from them. Long-term cultures established from chorionic villi appear to be no different from somatic cells with respect to X-chromosome inactivation.

Animals↗