Trophoblast retrieval and isolation from the cervix (TRIC) for noninvasive prenatal screening at 5 to 20 weeks of gestation.
OBJECTIVE
To use trophoblast cells accumulating in the endocervical canal at the beginning of pregnancy for noninvasive prenatal testing.


METHODS
Prospective, double-blinded test for fetal gender.


METHODS
Academic medical center.


METHODS
Fifty-six women with singleton pregnancies at gestational age 5-20 weeks.


METHODS
Isolation of fetal cells from resident maternal cells in endocervical specimens using anti-human leukocyte antigen G coupled to magnetic nanoparticles; cell phenotyping immunofluorescently with a panel of trophoblast subtype-specific proteins; DNA integrity assessment with terminal dUTP nick-end labeling (TUNEL); and polymerase chain reaction (PCR) and fluorescent in situ hybridization (FISH) to detect sex chromosomes in individual cells.


METHODS
Trophoblast phenotype, TUNEL index, and percentage male cells.


RESULTS
The women were given a routine Papanicolaou test; fetal genders were verified from medical records. Recovery after immunomagnetic isolation averaged 746±59 cells across gestational age, with 99% expressing chorionic gonadotropin, whereas the depleted cell fraction expressed none. The isolated cells had an extravillous trophoblast phenotype and intact nuclear DNA (>95%). Fetal gender was determined in 20 specimens without error by PCR. The FISH analysis of isolated cells from male specimens validated their fetal origin.


CONCLUSIONS
Noninvasive prenatal testing is feasible beginning at a gestational age of 5 weeks.
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan.
Jay M
JM
Bolnick
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan.
Brian A
BA
Kilburn
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan.
Swati
S
Bajpayee
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan.
Nitya
N
Reddy
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan.
Roohi
R
Jeelani
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan.
Barbara
B
Crone
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan.
Neil
N
Simmerman
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan.
Manivinder
M
Singh
Department of Obstetrics and Gynecology, Georgia Regents University, Augusta, Georgia.
Michael P
MP
Diamond
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan; Anatomy and Cell Biology, C.S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan; Program in Reproductive and Adult Endocrinology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland. Electronic address: d.armant@wayne.edu.
D Randall
DR
Armant
D000046:Academic Medical Centers; D015415:Biomarkers / Q000378:metabolism; D019070:Cell Lineage; D002478:Cells, Cultured; D002584:Cervix Uteri / Q000378:metabolism*; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000378:metabolism; D041321:Chromosomes, Human, X; D041322:Chromosomes, Human, Y; D053938:DNA Fragmentation; D004311:Double-Blind Method; D005260:Female; D005820:Genetic Testing; D005838:Genotype; D005865:Gestational Age; D006801:Humans; D018189:Immunomagnetic Separation; D017404:In Situ Hybridization, Fluorescence; D020287:In Situ Nick-End Labeling; D008297:Male; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D011446:Prospective Studies; D015203:Reproducibility of Results; D012732:Sex Determination Analysis; D014327:Trophoblasts / Q000378:metabolism*
D015415:Biomarkers; D018997:Chorionic Gonadotropin, beta Subunit, Human
Endocervical canal; fetal gender; prenatal testing; single-cell analysis; trophoblast