Trophoblast retrieval and isolation from the cervix (TRIC) for noninvasive prenatal screening at 5 to 20 weeks of gestation

Bolnick, Jay M.; Kilburn, Brian A.; Bajpayee, Swati; Reddy, N Ravindra; Jeelani, Roohi; Crone, Barbara; Simmerman, Neil; Singh, M.; Diamond, Michael P.; Armant, D. Randall

doi:10.1016/j.fertnstert.2014.04.008

Cited by 44 publications

(89 citation statements)

References 42 publications

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“…EVT cells are separated from maternal cells, using immunomagnetic isolation with antibody against human leukocyte antigen-G (HLA-G), an EVT-specific protein. The isolated cells have the expected EVT protein expression pattern (21). …”

Section: Introductionmentioning

confidence: 98%

“…Historically, the limiting step in employing these cells for placental evaluation was the inability to isolate them free of maternal cervical cells (20). Recently, trophoblast retrieval and isolation from the cervix (TRIC) has enabled isolation of EVT cells after a Papanicolaou procedure with 90–100% purity in a minimally invasive approach (21). EVT cells are separated from maternal cells, using immunomagnetic isolation with antibody against human leukocyte antigen-G (HLA-G), an EVT-specific protein.…”

Section: Introductionmentioning

confidence: 99%

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Noninvasive detection of trophoblast protein signatures linked to early pregnancy loss using trophoblast retrieval and isolation from the cervix (TRIC)

Fritz

Kohan-Ghadr

Bolnick

et al. 2015

Fertility and Sterility

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Objective To examine the expression pattern of biomarker proteins in extravillous trophoblast (EVT) cells obtained noninvasively by transcervical retrieval and isolation from the cervix (TRIC) in patients with early pregnancy loss, compared to control patients with uncomplicated term delivery. Design Case-control study. Setting Academic medical center. Patients Endocervical specimens were obtained from ongoing pregnancies at gestational ages of 5 to 10 weeks to generate an archive of EVT cells isolated by TRIC. Interventions Medical records were examined to select specimens from patients with either early pregnancy loss (EPL; N=10) or an uncomplicated term delivery (N=10), matched for gestational age at the time of endocervical sampling. Main Outcome Measures Known serum biomarkers for adverse pregnancy outcome that are expressed by EVT cells were evaluated by semi-quantitative immunocytochemistry, using antibodies against endoglin (ENG), FMS-like tyrosine kinase-1 (FLT-1), alpha-fetoprotein (AFP), pregnancy-associated plasma protein-A (PAPPA), galectin-13 (LGALS13), galectin-14 (LGALS14), and placental growth factor (PGF). Results EVT purity was over 95% in all specimens, based on chorionic gonadotropin expression; however, the number of EVT cells obtained was significantly lower in women with EPL than the control group. There was significant elevation of AFP, ENG, and FLT-1, and significant reduction of PAPP-A, LGALS14, and PGF in the EPL group, compared to controls. Conclusions In this pilot study, EVT cells isolated by TRIC early in gestation exhibit altered protein expression patterns prior to EPL, compared to uncomplicated term pregnancies.

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Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Noninvasive detection of trophoblast protein signatures linked to early pregnancy loss using trophoblast retrieval and isolation from the cervix (TRIC)

Fritz

Kohan-Ghadr

Bolnick

et al. 2015

Fertility and Sterility

Self Cite

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“…25 The nonbound cells were collected after magnetic immobilization of HLA-Gpositive trophoblast cells. A small aliquot of the isolated cell suspension was removed and counted to determine the number of cells recovered in the isolate.…”

Section: Immunomagnetic Isolation Of Trophoblast Cellsmentioning

confidence: 99%

“…The percentage of b-hCG-positive cells was quantified using secondary antibody conjugated to fluorescein isothiocyanate and a 4',6-diamidino-2-phenylindole (DAPI) nuclear counterstain, as described previously. 25 …”

Section: Immunomagnetic Isolation Of Trophoblast Cellsmentioning

confidence: 99%

“…[20][21][22] Intact fetal trophoblast cells are reliably obtained from ongoing pregnancies through safe, noninvasive endocervical collection, 23 preferably with a cytobrush and ThinPrep kit, 24 and can be separated from maternal cells using trophoblast retrieval and isolation from the cervix (TRIC). 25 Determination of fetal gender after TRIC is straight forward and was exceptionally accurate in 20 consecutive samples analyzed by polymerase chain reaction (PCR) as well as by fluorescence in situ hybridization (FISH). Examination of over 200 specimens collected between weeks 5 and 20 of pregnancy has shown TRIC to reliably provide 500 to 1000 trophoblast cells at purities above 95%, without interference by maternal obesity.…”

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confidence: 99%

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Trophoblast Retrieval and Isolation From the Cervix for Noninvasive, First Trimester, Fetal Gender Determination in a Carrier of Congenital Adrenal Hyperplasia

et al. 2016

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Congenital adrenal hyperplasia (CAH) is an autosomal recessive defect in cortisol biosynthesis that elevates fetal androgen levels to cause genital ambiguity and external genital masculinization in newborn females. Introducing dexamethasone in utero by 7 weeks gestation precludes virilization of affected females. However, identification of a male fetus prior to week 7 could avert the necessity of steroid treatment in half of pregnancies at risk of CAH. We recently introduced trophoblast retrieval and isolation from the cervix (TRIC), an approach that noninvasively isolate homogeneous trophoblast cells from pregnant women as early as 5 weeks gestation, using a Papanicolaou test. Here, we have used TRIC to correctly identify male fetal DNA when both parents were carriers of the mutation that produces CAH and previously produced an affected child. Trophoblast cells (1400) obtained by TRIC were assessed using immunocytochemistry with an antibody against the trophoblast-specific b subunit of human chorionic gonadotropin, which labeled 100% (17 of 17) of isolated cells, while none of the excluded maternal cervical cells were labeled. The isolated cells were examined by fluorescent in situ hybridization for chromosomes 18, X, and Y at a clinical cytogenetics laboratory, demonstrating 100% (18 of 18) of cells to be diploid 18/XY. Aliquots of DNA obtained from the isolated cells assayed for SRY and RNASEH genes by TaqMan assays confirmed a male fetus. This case study demonstrates the utility of TRIC to accurately identify fetal gender as a means of reducing the need for prophylactic administration of exogenous steroids in pregnancies at risk of CAH.

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Nondestructive Identification of Rare Trophoblastic Cells by Endoplasmic Reticulum Staining for Noninvasive Prenatal Testing of Monogenic Diseases

et al. 2020

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Noninvasive prenatal detection of monogenic diseases based on cell‐free DNA is hampered by challenges in obtaining a sufficient fraction and adequate quality of fetal DNA. Analyzing rare trophoblastic cells from Papanicolaou smears carrying the entire fetal genome provides an alternative method for noninvasive detection of monogenic diseases. However, intracellular labeling for identification of target cells can affect the quality of DNA in varying degrees. Here, a new approach is developed for nondestructive identification of rare fetal cells from abundant maternal cells based on endoplasmic reticulum staining and linear discriminant analysis (ER‐LDA). Compared with traditional methods, ER‐LDA has little effect on cell quality, allowing trophoblastic cells to be analyzed on the single‐cell level. Using ER‐LDA, high‐purity of trophoblastic cells can be identified and isolated at single cell resolution from 60 pregnancies between 4 and 38 weeks of gestation. Pathogenic variants, including –SEA/ deletion mutation and point mutations, in 11 fetuses at risk for α‐ or β‐thalassemia can be accurately detected by this test. The detection platform can also be extended to analyze the mutational profiles of other monogenic diseases. This simple, low‐cost, and noninvasive test can provide valuable fetal cells for fetal genotyping and holds promise for prenatal detection of monogenic diseases.

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Trophoblast retrieval and isolation from the cervix (TRIC) for noninvasive prenatal screening at 5 to 20 weeks of gestation

Cited by 44 publications

References 42 publications

Noninvasive detection of trophoblast protein signatures linked to early pregnancy loss using trophoblast retrieval and isolation from the cervix (TRIC)

Noninvasive detection of trophoblast protein signatures linked to early pregnancy loss using trophoblast retrieval and isolation from the cervix (TRIC)

Trophoblast Retrieval and Isolation From the Cervix for Noninvasive, First Trimester, Fetal Gender Determination in a Carrier of Congenital Adrenal Hyperplasia

Nondestructive Identification of Rare Trophoblastic Cells by Endoplasmic Reticulum Staining for Noninvasive Prenatal Testing of Monogenic Diseases

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