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> Anatomy > Embryonic Structure > Villi, Chorionic

Villi, Chorionic

Villi, Chorionic are the specialized projections that extend from the chorionic plate of the placenta.
They contain fetal blood vessels and are responsible for the transfer of nutrients, gases, and waste products between the maternal and fetal circulations.
Chorionic villi play a crucial role in the development and function of the placenta, enabling efficient exchange of essential substances for fetal growth and development.
Thier unique structure and dynamic interactions with the maternal environment make them a key area of study in reproductive biology and obstetrics.

Most cited protocols related to «Villi, Chorionic»

1
Derivation and Maintenance of Human Trophoblast Organoids
To obtain trophoblast-enriched cell suspensions, villi from first trimester placental tissue were sequentially digested with 0.2% trypsin-250 (Pan Biotech P10-025100P)/0.02% EDTA (Sigma #E9884)/PBS then collagenase V at 1.0mg/ml (Sigma #C9263) in Hams F12/10% FBS. Both digests were pooled, washed in Advanced DMEM/F12 medium (Gibco #12634-010) and re-suspended in ~10x volume growth-factor-reduced Matrigel (Corning #356231) on ice. 25µL drops were plated per well into a 48-well culture plate (Costar #3548), set at 37°C for 15min and overlaid with 250µL Trophoblast Organoid Medium (TOM, Supplementary Table 1b). Cultures were maintained in 5% CO2 in a humidified incubator at 37°C. Medium was replaced every 2-3d. Small organoid clusters became visible by ~7d and were passaged when at least 50% had reached a diameter of 200-300µm (usually between 7-10d). Mechanical disruption was achieved with Eppendorf Explorer Plus automatic pipettes on a mix cycle of 99 rounds (x4-5), maximum speed. Organoids from the same sample were initiated and maintained in the absence of each individual component to test its importance. Frozen stocks of organoids were made in 70% TOM, 20% FBS and 10% DMSO freeze medium and stored in liquid nitrogen. A step-by-step protocol of the derivation and maintenance of human trophoblast organoid cultures can be found at Nature Protocol Exchange24 .
Turco M.Y., Gardner L., Kay R.G., Hamilton R.S., Prater M., Hollinshead M., McWhinnie A., Esposito L., Fernando R., Skelton H., Reimann F., Gribble F., Sharkey A., Marsh S.G., O’Rahilly S., Hemberger M., Burton G.J, & Moffett A. (2018). Trophoblast organoids as a model for maternal-fetal interactions during human placentation. Nature, 564(7735), 263-267.
Publication 2018
ATF7IP protein, human Cells collagenase 1 Edetic Acid Freezing Growth Factor Homo sapiens matrigel Nitrogen Organoids PRSS2 protein, human Sulfoxide, Dimethyl Tissues TOMM70 protein, human Trophoblast Villi, Chorionic
2
Derivation and Maintenance of Human Trophoblast Organoids
To obtain trophoblast-enriched cell suspensions, villi from first trimester placental tissue were sequentially digested with 0.2% trypsin-250 (Pan Biotech P10-025100P)/0.02% EDTA (Sigma #E9884)/PBS then collagenase V at 1.0mg/ml (Sigma #C9263) in Hams F12/10% FBS. Both digests were pooled, washed in Advanced DMEM/F12 medium (Gibco #12634-010) and re-suspended in ~10x volume growth-factor-reduced Matrigel (Corning #356231) on ice. 25µL drops were plated per well into a 48-well culture plate (Costar #3548), set at 37°C for 15min and overlaid with 250µL Trophoblast Organoid Medium (TOM, Supplementary Table 1b). Cultures were maintained in 5% CO2 in a humidified incubator at 37°C. Medium was replaced every 2-3d. Small organoid clusters became visible by ~7d and were passaged when at least 50% had reached a diameter of 200-300µm (usually between 7-10d). Mechanical disruption was achieved with Eppendorf Explorer Plus automatic pipettes on a mix cycle of 99 rounds (x4-5), maximum speed. Organoids from the same sample were initiated and maintained in the absence of each individual component to test its importance. Frozen stocks of organoids were made in 70% TOM, 20% FBS and 10% DMSO freeze medium and stored in liquid nitrogen. A step-by-step protocol of the derivation and maintenance of human trophoblast organoid cultures can be found at Nature Protocol Exchange24 .
Turco M.Y., Gardner L., Kay R.G., Hamilton R.S., Prater M., Hollinshead M., McWhinnie A., Esposito L., Fernando R., Skelton H., Reimann F., Gribble F., Sharkey A., Marsh S.G., O’Rahilly S., Hemberger M., Burton G.J, & Moffett A. (2018). Trophoblast organoids as a model for maternal-fetal interactions during human placentation. Nature, 564(7735), 263-267.
Publication 2018
ATF7IP protein, human Cells collagenase 1 Edetic Acid Freezing Growth Factor Homo sapiens matrigel Nitrogen Organoids PRSS2 protein, human Sulfoxide, Dimethyl Tissues TOMM70 protein, human Trophoblast Villi, Chorionic
3
Prenatal Genetic Screening Protocol
This study was approved by the institutional review boards of the Chinese University of Hong Kong and Jinan University. Pregnant women requesting a prenatal diagnostic test referred to the two prenatal diagnosis centers were enrolled with written informed consent obtained from each participant. Prenatal samples including chorionic villi, amniotic fluid and cord blood were collected for DNA extraction and quality control (QC), while parental peripheral blood samples were collected whenever available either concurrently or after identification of a putative disease-associated variant for assessment of inheritance.
Genomic DNA was extracted with DNeasy Blood & Tissue Kit (cat. number/ID: 69506, Qiagen, Hilden, Germany) and treated with RNase (Qiagen, Hilden, Germany). DNA was subsequently quantified with the Qubit dsDNA HS Assay Kit (Invitrogen, Carlsbad, CA, USA) and DNA integrity was assessed by gel electrophoresis. Quantitative fluorescence polymerase chain reaction (QF-PCR) was conducted with 100 ng DNA and two panels of short tandem repeat (STR) markers (P1 and XY) located on chromosomes 18, X, and Y for exclusion of maternal cell admixture and polyploidy16 (link). Subsequently, after exclusion, each DNA sample was subjected for routine CMA and low-pass GS in parallel.
Wang H., Dong Z., Zhang R., Chau M.H., Yang Z., Tsang K.Y., Wong H.K., Gui B., Meng Z., Xiao K., Zhu X., Wang Y., Chen S., Leung T.Y., Cheung S.W., Kwok Y.K., Morton C.C., Zhu Y, & Choy K.W. (2019). Low-pass genome sequencing versus chromosomal microarray analysis: implementation in prenatal diagnosis. Genetics in medicine : official journal of the American College of Medical Genetics, 22(3), 500-510.
Publication 2019
Amniotic Fluid Antenatal Screenings Biological Assay BLOOD Chinese Chromosomes, Human, Pair 18 Diagnosis DNA, Double-Stranded Electrophoresis Endoribonucleases Ethics Committees, Research Fluorescence Genome Intrauterine Diagnoses Parent Pattern, Inheritance Polymerase Chain Reaction Pregnant Women Short Tandem Repeat Stem Cells Tests, Diagnostic Tissues Umbilical Cord Blood Villi, Chorionic
4
Placental DNA Methylation in Early-Onset Preeclampsia
Whole chorionic villi were sampled from placentas delivered at Women's Hospital in Vancouver, Canada. Clinical criteria for EOPET were defined using the Canadian guidelines (Magee et al., 2008 (link)). For DNA methylation analysis, samples were further sub-classified by the presence/absence of severe proteinuria (>3 g/day), coincident IUGR, gestational diabetes and HELLP syndrome, defined by hemolysis, elevated liver enzymes and low platelet count. IUGR was defined as (i) birthweight below the third percentile for gender and gestational age using Canadian population parameters or (ii) birthweight below the 10th percentile with persistent uterine artery notching at 22–25 weeks, absent or reversed end diastolic velocity on umbilical artery Doppler, or oligohydramnios. As gestational age is a large determinant of DNA methylation in the placenta (Novakovic et al., 2011 (link)), it is critical that placentas from pre-eclamptic pregnancies are compared with gestational age-matched controls (Yuen et al., 2010 (link)). We therefore compared with placentas obtained from chromosomally normal losses or births due to a mix of etiologies (premature rupture of membranes, loss of amniotic fluid and cervical incompetence) with no evidence of placental abnormality based on a pathological exam (Table I; Supplementary data, Table SI). The use of such pre-term samples is validated by our previous observation that the DNA methylation profiles of these premature-controls were more similar to term-controls than to the gestational age (GA) age-matched EOPET placentas (Yuen et al., 2010 (link)). Chorionic villi from at least two sites (center and perimeter) on the fetal side of the placenta were sampled, rinsed of maternal blood. DNA was extracted from each and combined in equal quantities for better representation of placenta-wide changes. It should be noted that four of the samples in this study have been previously assessed for methylation changes (using less-comprehensive microarrays) (Yuen et al., 2009 (link), 2010 (link)).

Clinical information of samples used in Illumina methylation array.

EOPET (n = 20) mean (range)CONTROL (n = 20) mean (range)P-value
Gestational age (weeks)31.8 (24.9–37.3)31.8 (25.0–37.3)0.931
Maternal age (years)33.5 (19.7–42.9)31.5 (22.2–38.7)0.289
Birthweight (g)1451 (440–3685)1940 (758–3470)0.052
Blair J.D., Yuen R.K., Lim B.K., McFadden D.E., von Dadelszen P, & Robinson W.P. (2013). Widespread DNA hypomethylation at gene enhancer regions in placentas associated with early-onset pre-eclampsia. Molecular Human Reproduction, 19(10), 697-708.
Publication 2013
Amniotic Fluid Birth Weight BLOOD Diastole DNA Methylation Enzymes Fetal Growth Retardation Fetal Membranes, Premature Rupture Fetus Gestational Age Gestational Diabetes HELLP Syndrome Hemolysis Liver Methylation Microarray Analysis Mothers Oligohydramnios Perimetry Placenta Platelet Counts, Blood Pre-Eclampsia Pregnancy Premature Birth Umbilical Arteries Uterine Arteries Uterine Cervical Incompetence Villi, Chorionic
5
Trophoblast Organoid Differentiation
Trophoblast organoids were passaged and plated into 35mm dishes or ibidi μ-dishes (Thermo Scientific #81156). Differentiation was achieved through a modification of the protocol used by Okae et al18 (link). After passaging, organoids were maintained in TOM for 3-4d and switched to EVT medium (EVTM: advanced DMEM/F12, 0.1mM 2-mercaptoethanol (Gibco #31350), 0.5% Penicillin-streptomycin, 0.3% BSA (Sigma #A8412), 1% ITS-X supplement (Gibco #51500-056), 100ng/mL NRG1 (Cell Signaling #5218SC), 7.5μM A83-01 (Tocris Biotechne #2939), 4% Knockout serum replacement (ThermoFisher #10828010). When organoids showed outgrowth of cells (typically 7-10d), the medium was changed to EVTM without NRG1 for a further 7-10d. For comparison, fresh placental villi were embedded into 300uL Matrigel in ibidi μ-dishes and grown under the same conditions.
Turco M.Y., Gardner L., Kay R.G., Hamilton R.S., Prater M., Hollinshead M., McWhinnie A., Esposito L., Fernando R., Skelton H., Reimann F., Gribble F., Sharkey A., Marsh S.G., O’Rahilly S., Hemberger M., Burton G.J, & Moffett A. (2018). Trophoblast organoids as a model for maternal-fetal interactions during human placentation. Nature, 564(7735), 263-267.
Publication 2018
2-Mercaptoethanol A-83-01 Cells Dietary Supplements Hyperostosis, Diffuse Idiopathic Skeletal matrigel NRG1 protein, human Organoids Penicillins Serum Streptomycin Trophoblast Villi, Chorionic

Most recents protocols related to «Villi, Chorionic»

1
Maternal-Placental miRNA Expression Dynamics
To distinguish between maternal and placental involvement in miRNA expression, we exposed primary cell cultures from early stage human placenta villus trophoblast (HVT) to EV pellets that were obtained from the HP and GVC groups. The primary ECs representing the maternal side were also exposed to EVs of NP, as well as EVs of HP and women with GVC.
Aharon A., Rebibo-Sabbah A., Ahmad R.S., Dangot A., Bar-Lev T.H., Brenner B., Cohen A.H., David C.B., Weiner Z, & Solt I. (2023). Associations of maternal and placental extracellular vesicle miRNA with preeclampsia. Frontiers in Cell and Developmental Biology, 11, 1080419.
Full text: Click here
Publication 2023
Cells Homo sapiens MicroRNAs Pellets, Drug Placenta Trophoblast Villi, Chorionic Woman
2
Establishing Cholestatic Cell Model
The human chorionic villus trophoblast cell line (HTR8/SVneo cells) was obtained from the American Type Culture Center (ATCC) and identified by STR Authentication. HTR8/Svneo cells were cultured in 90% RPMI 1640 medium, 1.0% dual antibodies (penicillin and streptomycin), and 10% fetal bovine serum in a complete medium. HTR8/Svneo cells were seeded in sterile 6-well plates and cultured until the cell density reached 70%, and then treated with 100 μmol/L sodium taurocholate (TCA) for 24 h to establish a cholestatic cell model [23 (link)].
Kong Y., Zhan Y., Chen D., Deng X., Liu X., Xu T, & Wang X. (2023). Unique microRNA expression profiles in plasmic exosomes from intrahepatic cholestasis of pregnancy. BMC Pregnancy and Childbirth, 23, 147.
Full text: Click here
Publication 2023
Antibodies Cell Lines Cells Central American People Cholestasis Cultured Cells Fetal Bovine Serum Homo sapiens Penicillins Sterility, Reproductive Streptomycin Taurocholic Acid, Monosodium Salt Trophoblast Villi, Chorionic
3
Chromosomal Abnormalities in Miscarriage Samples
All fresh miscarriage specimens were rinsed with saline solution for three times. Chorionic villi were separated from maternal decidua according to the standardized technology [9 (link)]. Samples where chorionic villi could not be clearly identified were excluded from this study. Genomic DNA was extracted from chorionic villi with the protocol of QIAamp DNA Mini Kit (Qiagen, Germany). Chromosomal abnormalities of POCs were detected by two CMA platforms in the current study, including CytoScan 750K array (Affymetrix, USA) and HumanCyto12-SNP array (Illumina, USA). SNP array experiments and molecular karyotype analysis for both platforms were performed as previously described [10 (link)]. Quantitative fluorescent polymerase chain reaction (QF-PCR) was subsequently performed to identify the percentage of maternal and foetal DNA if MCC was detected by CMA. Significant MCC referred to the proportion of MCC exceeding 30%. Samples with significant MCC were excluded from our study.
The two platforms could detect CNVs at an effective minimal resolution of 100 kb and regions of allelic homozygosity (ROHs) at a threshold of 5 Mb. Mosaicism for aneuploidies or CNVs ≥ 5 Mb was reported when the detection threshold of 30% was exceeded. CNVs were further classified as partial aneuploidy (CNVs ≥ 10 Mb, large CNVs) and microdeletions/microduplications (CNVs < 10 Mb, submicroscopic CNVs) based on their sizes. Pathogenicity of detected CNVs were evaluated according to the American College of Medical Genetics and Genomics (ACMG) and the Clinical Genome Resource (ClinGen) [11 (link)].
Xia Z., Zhou R., Li Y., Meng L., Huang M., Tan J., Qiao F., Zhu H., Hu P., Zhu Q., Xu Z, & Wang Y. (2023). Reproductive outcomes in couples with sporadic miscarriage after embryonic chromosomal microarray analysis. Annals of Medicine, 55(1), 837-848.
Publication 2023
Alleles Aneuploidy Care, Prenatal Chromosome Aberrations Decidua Genome Homozygote Karyotype Mosaicism Pathogenicity Polymerase Chain Reaction Saline Solution Spontaneous Abortion Villi, Chorionic
4
Placental Tissue Explants for in vitro Study
Human third-trimester placentas (36 to 40 weeks of pregnancy), n = 4, were collected after elective cesarean deliveries at the Clinics Hospital of the Federal University of Uberlândia (HC-UFU), MG, Brazil. Placental tissues were collected based on exclusion criteria, as follows: pre-eclampsia, chronic hypertension, infectious diseases including toxoplasmosis, chorioamnionitis, chronic kidney disease, heart disease, connective tissue disease, pre-existing diabetes mellitus, gestational diabetes mellitus and other pathological manifestations. Briefly, placental tissues were washed in sterile PBS to remove excess blood, then aseptically dissected to remove endometrial tissue and fetal membranes up to 1 h after collection. Terminal chorionic villi containing five to seven free tips per explant were harvested and added to 96-well microplates (one villus per well) in 200 µL/well of fresh RPMI 1640 medium supplemented with 100 U/mL penicillin, 100 µg/mL streptomycin and 10% FBS for 24 h at 37°C in a humidified atmosphere containing CO2 (5%) (Silva et al., 2017 (link)).
Martínez A.F., Teixeira S.C., de Souza G., Rosini A.M., Júnior J.P., Melo G.N., Blandón K.O., Gomes A.O., Ambrósio S.R., Veneziani R.C., Bastos J.K., Martins C.H., Ferro E.A, & Barbosa B.F. (2023). Leaf hydroalcoholic extract and oleoresin from Copaifera multijuga control Toxoplasma gondii infection in human trophoblast cells and placental explants from third-trimester pregnancy. Frontiers in Cellular and Infection Microbiology, 13, 1113896.
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Publication 2023
Atmosphere BLOOD Cesarean Section Chorioamnionitis Chronic Kidney Diseases Communicable Diseases Connective Tissue Diseases Endometrium Fetal Membranes Gestational Diabetes Heart Diseases High Blood Pressures Homo sapiens Penicillins Placenta Pre-Eclampsia Pregnancy States, Prediabetic Sterility, Reproductive Streptomycin Tissues Toxoplasmosis Villi, Chorionic
5
Immunohistochemical Analysis of Placental Proteins
All tissue specimens were formalin-fixed and embedded in paraffin, and 4-µm thick sections were obtained on silanized glass slides. The slides were dewaxed in xylene and hydrated through a graded series of ethanol. Endogenous peroxidase was blocked with 3% hydrogen peroxide. Antigen retrieval for IFNA4, IRF3, and RIG-I analysis was performed by incubation of the slides in 50 mM Tris-ethylenediaminetetraacetic acid (EDTA) buffer, pH 9.0 (code 52367, Dako, Carpinteria, CA, USA), in a water bath for 20 min at 95 °C, followed by washing in running water and distilled water for 5 min each. Then, they were incubated overnight at 4 °C in the presence of the primary antibody, polyclonal rabbit anti-IFNA4 primary antibody (1:300, ab232899, Abcam, Boston, MA, USA), monoclonal rabbit anti-IRF3 (1:60, ab76409, Abcam), or polyclonal goat anti-RIG-I/DDX58 (1:30, ab111037, Abcam). The specific antigen–antibody reaction for IFNA4 was detected with the NovoLink Polymer Detection System Kit (code RE7280-CE, Leica Microsystems Inc., Newcastle Upon Tyne, UK), IRF3 was detected with the Detection Kit-Micro-polymer (ab236466, Abcam, Boston, MA, USA), and RIG-I/DDX8 was detected with the Stain MAX PO Universal Immuno-peroxidase Polymer (Histofine 414161F, Nichirei Biosciences Inc., Tokyo, JPN), according to the manufacturer’s instructions. The reactions were visualized using the 3,3′-diaminobenzidine-tetrahydrochloride (DAB) chromogen (Sigma) and counterstained with Carazzi hematoxylin. All reactions were accompanied by both positive (placenta and tonsil tissue) and negative controls. For the negative control of nonspecific binding, the primary antibodies were replaced with normal serum, under the same experimental conditions. For each protein analysis, we used five random regions on the slides for the placental villus and three regions for the decidua. The Image-Pro plus version 4.5.0.29 software was used for quantification, by counting the positive area or optical density intensity (ODI) by the total area or length of the basement membrane, for the placental decidua. For normalization, a 10× ocular lens was used for the decidua and 20× for the placental villus.
Branco A.C., De Oliveira E.A., Pereira N.Z., Alberca R.W., Duarte-Neto A.N., Da Silva L.F., Luiz F.G., Pereira N.V., Sotto M.N., Dejani N.N., Rondó P.H., Avvad-Portari E., De Vasconcelos Z.F., Duarte A.J., Azamor T, & Sato M.N. (2023). Obesity Induces an Impaired Placental Antiviral Immune Response in Pregnant Women Infected with Zika Virus. Viruses, 15(2), 320.
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Publication 2023
Antibodies Antibodies, Anti-Idiotypic Antigen-Antibody Reactions Antigens azo rubin S Bath DDX58 protein, human Decidua Edetic Acid Ethanol Formalin Goat Hematoxylin Immunoglobulins Immunoperoxidase Techniques IRF3 protein, human Lens, Crystalline Membrane, Basement Palatine Tonsil Paraffin Embedding Peroxidase Peroxides Placenta Polymers Proteins Rabbits Serum Stains Tissues Tritium Tromethamine Villi, Chorionic Vision Xylene

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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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More about "Villi, Chorionic"

Chorionic villi, the specialized projections extending from the chorionic plate of the placenta, play a crucial role in fetal development.
These villi contain fetal blood vessels and facilitate the efficient transfer of nutrients, gases, and waste products between the maternal and fetal circulations.
The unique structure and dynamic interactions of chorionic villi with the maternal environment make them a key area of study in reproductive biology and obstetrics.
The chorionic villi are composed of fetal mesenchymal and trophoblastic cells, and their development is essential for the proper functioning of the placenta.
The trophoblast cells, which form the outer layer of the villi, are responsible for the invasion and remodeling of the maternal uterine tissues, enabling the establishment of the uteroplacental circulation.
Researchers often utilize various techniques and materials to study chorionic villi, such as the FBS (Fetal Bovine Serum) and DMEM (Dulbecco's Modified Eagle Medium) for cell culture, the QIAamp DNA Mini Kit and DNeasy Blood and Tissue Kit for DNA extraction, and Trypsin for cell dissociation.
Additionally, the use of Penicillin/Streptomycin, DMEM/F12, and RPMI 1640 medium can help maintain the viability and growth of chorionic villi cells in vitro.
The Matrigel substrate, a complex extracellular matrix derived from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells, can also be employed to mimic the three-dimensional (3D) environment of the placenta and study the behavior and differentiation of chorionic villi cells.
Understanding the structure, function, and development of chorionic villi is crucial for advancing our knowledge of placental biology, fetal growth, and the identification of potential biomarkers or therapeutic targets related to pregnancy-related disorders, such as preeclampsia, intrauterine growth restriction, and gestational diabetes.