Immunocytochemical identification of cytotrophoblast from other mononuclear cell populations isolated from first-trimester human chorionic villi

1985 ◽  
Vol 76 (1) ◽  
pp. 189-197
Author(s):  
B.H. Butterworth ◽  
Y.W. Loke
Keyword(s):  
Monoclonal Antibodies ◽  
Mononuclear Cell ◽  
Chorionic Villus ◽  
Cell Types ◽  
First Trimester ◽  
Cell Populations ◽  
Double Labelling ◽  
Chorionic Villi ◽  
Isolation Methods ◽  
Human Placental Cells

Trophoblast biologists are often uncertain as to what cell types they are investigating because the mononuclear cell populations prepared from trypsinization of human first-trimester chorionic villi are morphologically very similar. In the present study, immunocytochemical and phagocytic markers have been used to distinguish cytotrophoblast populations from cell types derived from the mesenchyme of the chorionic villus. Two anti-trophoblast monoclonal antibodies generated in our own laboratory (18B/A5 and 18A/C4) were found to be very efficient in identifying cytotrophoblast, which made up 35–40% of the cells in a smear. Most cytotrophoblast cells did not stain with a monoclonal anti-HLA-A,B,C antibody but a few cells (5%) were found to express both trophoblast and HLA-A,B,C antigens by a double-labelling technique. Endothelial cells from villous capillaries could be identified by a rabbit anti-factor VIII antibody. These cells formed 28% of the population in a cytospin smear. Macrophages from the villous mesenchyme were less readily separable as neither specific monoclonal antibodies nor localization of enzymes were found to be effective. However, these cells could be identified by their ability to phagocytose carmine. About 15% of the cells in a smear consisted of macrophages. The procedure described should prove useful in judging the efficiency of isolation methods from human placental cells.

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

1984 ◽  
Vol 30 (12) ◽  
pp. 2040-2042 ◽  
Author(s):  
P L Trnka ◽  
E Pergament ◽  
N G Anderson
Keyword(s):  
Cell Types ◽  
First Trimester ◽  
Gene Products ◽  
Two Dimensional ◽  
Chorionic Villi ◽  
Two Dimensional Electrophoresis ◽  
Chorionic Plate ◽  
Reference Map ◽  
Dimensional Electrophoresis ◽  
Fetal Genotype

Abstract 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.

Effects of hypoxia on heme oxygenase expression in human chorionic villi explants and immortalized trophoblast cells

2003 ◽  
Vol 284 (3) ◽  
pp. H853-H858 ◽  
Author(s):  
S. D. Appleton ◽  
G. S. Marks ◽  
K. Nakatsu ◽  
J. F. Brien ◽  
G. N. Smith ◽  
...  
Keyword(s):  
Protein Expression ◽  
Protein Content ◽  
Heme Oxygenase ◽  
Protein Function ◽  
Cell Types ◽  
First Trimester ◽  
Trophoblast Cells ◽  
Human Trophoblast ◽  
Chorionic Villi ◽  
Enzymatic Function

Although hypoxia induces heme oxygenase (HO)-1 protein and mRNA expression in many cell types, hypoxia has also been shown to decrease HO-1 mRNA and protein expression. We tested the hypothesis that 24-h preexposure to hypoxia in human placental preparations suppresses HO protein expression and enzymatic function. Immortalized HTR-8/SVneo first-trimester trophoblast cells and explants of normal human chorionic villi (CV) from term placentas were cultured for 24 h in 1%, 5%, or 20% O2. HO protein levels were determined by Western blot analysis, and microsomal HO activity was measured. HO-2 protein content was decreased by 17% and 5% in human trophoblast cells after 24-h exposure to 1% and 5% O2, respectively, versus 20% O2. In contrast, HO-2 protein content in CV explants was unaffected by changes in oxygenation. HO-1 protein content, which was barely detectable in both biological systems, was not affected by changes in oxygenation. Similarly, HO enzymatic activity was unchanged in both preparations after 24-h exposure to 1%, 5%, or 20% O2. The above data do not support the hypothesis that hypoxia in the human placenta suppresses both HO protein content and HO protein function. The present observations reinforce the necessity to determine both HO protein expression and function.

scholarly journals Cell-specific characterization of the placental methylome

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Victor Yuan ◽  
Desmond Hui ◽  
Yifan Yin ◽  
Maria S. Peñaherrera ◽  
Alexander G. Beristain ◽  
...  
Keyword(s):  
Placental Tissue ◽  
Cell Types ◽  
First Trimester ◽  
R Package ◽  
Cell Populations ◽  
Placental Development ◽  
Methylation Array ◽  
Cell Composition ◽  
Distinct Cell ◽  
Hofbauer Cells

Abstract Background DNA methylation (DNAm) profiling has emerged as a powerful tool for characterizing the placental methylome. However, previous studies have focused primarily on whole placental tissue, which is a mixture of epigenetically distinct cell populations. Here, we present the first methylome-wide analysis of first trimester (n = 9) and term (n = 19) human placental samples of four cell populations: trophoblasts, Hofbauer cells, endothelial cells, and stromal cells, using the Illumina EPIC methylation array, which quantifies DNAm at > 850,000 CpGs. Results The most distinct DNAm profiles were those of placental trophoblasts, which are central to many pregnancy-essential functions, and Hofbauer cells, which are a rare fetal-derived macrophage population. Cell-specific DNAm occurs at functionally-relevant genes, including genes associated with placental development and preeclampsia. Known placental-specific methylation marks, such as those associated with genomic imprinting, repetitive element hypomethylation, and placental partially methylated domains, were found to be more pronounced in trophoblasts and often absent in Hofbauer cells. Lastly, we characterize the cell composition and cell-specific DNAm dynamics across gestation. Conclusions Our results provide a comprehensive analysis of DNAm in human placental cell types from first trimester and term pregnancies. This data will serve as a useful DNAm reference for future placental studies, and we provide access to this data via download from GEO (GSE159526), through interactive exploration from the web browser (https://robinsonlab.shinyapps.io/Placental_Methylome_Browser/), and through the R package planet, which allows estimation of cell composition directly from placental DNAm data.

scholarly journals INVESTIGATION OF TROPHOBLAST IN TORCH INFECTION ON ABORTION MATERIAL IN THE FIRST TRIMESTER OF PREGNANCY

2019 ◽  
Vol 18 (4) ◽  
pp. 58-62
Author(s):  
A. V. Goshovskaya ◽  
V. M. Goshovsky ◽  
S. M. Yasnikovska
Keyword(s):  
Gestational Age ◽  
Imaging System ◽  
Chorionic Villus ◽  
First Trimester ◽  
Control Group ◽  
Chorionic Villi ◽  
Infectious Process ◽  
First Trimester Of Pregnancy ◽  
Torch Infection ◽  
Immunohistochemical Studies

This study is a fragment of a series of immunohistochemical studies of trophoblast with TORCH infection, which are scheduled to be carried out at different gestational dates. This article deals with the results of trophoblast studies at the gestational age of 7-8 weeks. The study examined abortion material 7-8 weeks of gestation. The main group of the study consisted of 18 observations of TORCH infection, and the control group consisted of 17 observations of an aborted pregnancy without signs of an infectious process (abortion for social reasons). An immunohistochemical procedure was performed on metalloproteinases-2 with primary antibodies and polymer antigen imaging system using DAKO diaminobenzidine. The method of computer microdensitometry in a specialized computer program ImageJ evaluated the optical density of the color. According to the results of immunohistochemical studies using computer microdensitometry at the gestational age of 7-8 weeks, both with TORCH infection and without it an infectious process, strong expression of metalloproteinase-2 is observed in the invasive trophoblast, the smallest – in the syncytotrophoblast of the chorionic villi, and intermediate values are noted in chorionic villus cytotrophoblast and cell column cytotrophoblast. With TORCH infection the expression of metalloproteinase-2 decreases in all the four types of trophoblast (cytotrophoblast of chorionic villi; cytotrophoblast of cell columns; invasive cytotrophoblast in endometrial fragments), except for syncytotrophoblast of chorionic villi was found.

scholarly journals Cell-specific Characterization of the Placental Methylome

2020 ◽  
Author(s):  
Victor Yuan ◽  
Desmond Hui ◽  
Yifan Yin ◽  
Maria Peñaherrera ◽  
Alexander Beristain ◽  
...  
Keyword(s):  
Placental Tissue ◽  
Cell Types ◽  
First Trimester ◽  
R Package ◽  
Cell Populations ◽  
Placental Development ◽  
Methylation Array ◽  
Cell Composition ◽  
Distinct Cell ◽  
Hofbauer Cells

Abstract Background: DNA methylation (DNAm) profiling has emerged as a powerful tool for characterizing the placental methylome. However, previous studies have focused primarily on whole placental tissue, which is a mixture of epigenetically distinct cell populations. Here, we present the first methylome-wide analysis of first trimester (n=9) and term (n=19) human placental samples of four cell populations: trophoblasts, Hofbauer cells, endothelial cells, and stromal cells, using the Illumina EPIC methylation array, which quantifies DNAm at >850,000 CpGs.Results: The most distinct DNAm profiles were those of placental trophoblasts, which are central to many pregnancy-essential functions, and Hofbauer cells, which are a rare fetal-derived macrophage population. Cell-specific DNAm occurs at functionally-relevant genes, including genes associated with placental development and preeclampsia. Known placental-specific methylation marks, such as those associated with genomic imprinting, repetitive element hypomethylation, and placental partially methylated domains, were found to be more pronounced in trophoblasts and often absent in Hofbauer cells. Lastly, we characterize the cell composition and cell-specific DNAm dynamics across gestation.Conclusions: Our results provide a comprehensive analysis of DNAm in human placental cell types from first trimester and term pregnancies. This data will serve as a useful DNAm reference for future placental studies, and we provide access to this data via download from GEO (GSE159526), through interactive exploration from the web browser (https://robinsonlab.shinyapps.io/Placental_Methylome_Browser/), and through the R package planet, which allows estimation of cell composition directly from placental DNAm data.

scholarly journals Cell-specific Characterization of the Placental Methylome

2020 ◽  
Author(s):  
Victor Yuan ◽  
Desmond Hui ◽  
Yifan Yin ◽  
Maria Peñaherrera ◽  
Alexander Beristain ◽  
...  
Keyword(s):  
Placental Tissue ◽  
Cell Types ◽  
First Trimester ◽  
R Package ◽  
Cell Populations ◽  
Placental Development ◽  
Methylation Array ◽  
Cell Composition ◽  
Distinct Cell ◽  
Hofbauer Cells

Abstract Background: DNA methylation (DNAm) profiling has emerged as a powerful tool for characterizing the placental methylome. However, previous studies have focused primarily on whole placental tissue, which is a mixture of epigenetically distinct cell populations. Here, we present the first methylome-wide analysis of first trimester (n=9) and term (n=19) human placental samples of four cell populations: trophoblasts, Hofbauer cells, endothelial cells, and stromal cells, using the Illumina EPIC methylation array, which quantifies DNAm at >850,000 CpGs.Results: The most distinct DNAm profiles were those of placental trophoblasts, which are central to many pregnancy-essential functions, and Hofbauer cells, which are a rare fetal-derived macrophage population. Cell-specific DNAm occurs at functionally-relevant genes, including genes associated with placental development and preeclampsia. Known placental-specific methylation marks, such as those associated with genomic imprinting, repetitive element hypomethylation, and placental partially methylated domains, were found to be more pronounced in trophoblasts and often absent in Hofbauer cells. Lastly, we characterize the cell composition and cell-specific DNAm dynamics across gestation.Conclusions: Our results provide a comprehensive analysis of DNAm in human placental cell types from first trimester and term pregnancies. This data will serve as a useful DNAm reference for future placental studies, and we provide access to this data via download from dbGAP (phs002013.v1.p1), through interactive exploration from the web browser (https://robinsonlab.shinyapps.io/Placental_Methylome_Browser/), and through the R package planet, which allows estimation of cell composition directly from placental DNAm data.

scholarly journals Features of the histological structure of trophoblast and chorionic villi with recurrent pregnancy loss in women with thrombophilias

2017 ◽  
Vol 25 (4) ◽  
pp. 621-641
Author(s):  
A. I. Mirov ◽  
O. N. Kharkevich ◽  
O. E. Golofast ◽  
I. B. Glukhovets
Keyword(s):  
Significant Role ◽  
Pregnancy Loss ◽  
Recurrent Pregnancy Loss ◽  
Chorionic Villus ◽  
Fetal Loss ◽  
First Trimester ◽  
Control Group ◽  
Histological Structure ◽  
Chorionic Villi ◽  
First Trimester Of Pregnancy

The frequency of recurrent pregnancy loss does not tend to decrease. This pathology continues to be one of the important problems of modern medicine. It is known that thrombophilia can play a significant role in the etiology of spontaneous reproductive losses. However, the pathogenesis of recurrent spontaneous loss of pregnancy in the presence of maternal thrombophilia is not fully understood. Aim. To identify the features of the histological structure of trophoblasts and chorionic villi in the first trimester of pregnancy in women with thrombophilia and recurrent pregnancy loss, with careful exclusion of other possible causes of fetal loss syndrome. Material and Methods. Histological examination of 49 chorion tissue samples from 24 patients with thrombophilia and recurrent pregnancy loss in the first trimester (study group) was performed. The controls were samples of chorion tissue taken during artificial abortion in 33 healthy women who had a history of 2 or more spontaneous labor without significant complications. Thrombophilia diagnosis and hemostasis system state evaluation was performed for all patients on the basis of analysis of 30 parameters according to standard methods. All studies were conducted at the Regional clinical hospital № 8 in Ryazan as well as the scientific and clinical center of hematology, oncology and immunology of the Ryazan State Medical University named after academician I.P. Pavlov of Health Ministry of the Russian Federation. Statistical processing of the obtained results was carried out with the help of computer program package Statistica (version 10). Results. Significant differences in the histological structure of trophoblast and chorionic villi in the studied women were revealed, in comparison with those in the control group. It is proved that the presence of thrombophilia negatively affects the process of embryogenesis and contributes to a significant reduction in the area of the chorionic villus vessels in the first trimester of pregnancy. Conclusion. It is proved that the presence of thrombophilia has a negative effect on the process of embryogenesis and significantly reduces the vascular area of chorionic villi that can probably play a significant role in the pathogenesis of recurrent pregnancy loss.

scholarly journals Cell-specific Characterization of the Placental Methylome

2020 ◽  
Author(s):  
Victor Yuan ◽  
Desmond Hui ◽  
Yifan Yin ◽  
Maria Peñaherrera ◽  
Alexander Beristain ◽  
...  
Keyword(s):  
Placental Tissue ◽  
Cell Types ◽  
First Trimester ◽  
R Package ◽  
Cell Populations ◽  
Placental Development ◽  
Methylation Array ◽  
Cell Composition ◽  
Distinct Cell ◽  
Hofbauer Cells

Abstract Background DNA methylation (DNAm) profiling has emerged as a powerful tool for characterizing the placental methylome. However, previous studies have focused primarily on whole placental tissue, which is a mixture of epigenetically distinct cell populations. Here, we present the first methylome-wide analysis of first trimester (n = 9) and term (n = 19) human placental samples of four cell populations: trophoblasts, Hofbauer cells, endothelial cells, and stromal cells, using the Illumina EPIC methylation array, which quantifies DNAm at > 850,000 CpGs. Results The most distinct DNAm profiles were those of placental trophoblasts, which are central to many pregnancy-essential functions, and Hofbauer cells, which are a rare understudied macrophage population thought to derive from fetal monocytes. Cell-specific DNAm occurs at functionally-relevant genes, including genes associated with placental development and preeclampsia. Known placental-specific methylation marks, such as those associated with genomic imprinting, repetitive element hypomethylation, and placental partially methylated domains, were found to be more pronounced in trophoblasts and often absent in Hofbauer cells. Lastly, we characterize the cell composition and cell-specific DNAm dynamics across gestation. Conclusions Our results provide a comprehensive analysis of DNAm in human placental cell types from first trimester and term pregnancies. This data will serve as a useful DNAm reference for future placental studies, and we provide access to this data via download from dbGAP (phs002013.v1.p1), through interactive exploration from the web browser (https://robinsonlab.shinyapps.io/Placental_Methylome_Browser/), and through the R package planet, which allows estimation of cell composition directly from placental DNAm data.

scholarly journals Single-cell transcriptomic analysis of mIHC images via antigen mapping

2021 ◽  
Vol 7 (10) ◽  
pp. eabc5464
Author(s):  
Kiya W. Govek ◽  
Emma C. Troisi ◽  
Zhen Miao ◽  
Rachael G. Aubin ◽  
Steven Woodhouse ◽  
...  
Keyword(s):  
Single Cell ◽  
Spatial Patterns ◽  
Cell Types ◽  
Level Of Detail ◽  
Cell Populations ◽  
Sequencing Data ◽  
Spatially Resolved ◽  
Murine Spleen ◽  
Single Cell Rna Sequencing ◽  
Antibody Panel

Highly multiplexed immunohistochemistry (mIHC) enables the staining and quantification of dozens of antigens in a tissue section with single-cell resolution. However, annotating cell populations that differ little in the profiled antigens or for which the antibody panel does not include specific markers is challenging. To overcome this obstacle, we have developed an approach for enriching mIHC images with single-cell RNA sequencing data, building upon recent experimental procedures for augmenting single-cell transcriptomes with concurrent antigen measurements. Spatially-resolved Transcriptomics via Epitope Anchoring (STvEA) performs transcriptome-guided annotation of highly multiplexed cytometry datasets. It increases the level of detail in histological analyses by enabling the systematic annotation of nuanced cell populations, spatial patterns of transcription, and interactions between cell types. We demonstrate the utility of STvEA by uncovering the architecture of poorly characterized cell types in the murine spleen using published cytometry and mIHC data of this organ.

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