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

Fertil Steril. 2014 Jul;102(1):135-142.e6. doi: 10.1016/j.fertnstert.2014.04.008. Epub 2014 May 10.

Abstract

Objective: To use trophoblast cells accumulating in the endocervical canal at the beginning of pregnancy for noninvasive prenatal testing.

Design: Prospective, double-blinded test for fetal gender.

Setting: Academic medical center.

Patient(s): Fifty-six women with singleton pregnancies at gestational age 5-20 weeks.

Intervention(s): 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.

Main outcome measure(s): Trophoblast phenotype, TUNEL index, and percentage male cells.

Result(s): 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.

Conclusion(s): Noninvasive prenatal testing is feasible beginning at a gestational age of 5 weeks.

Keywords: Endocervical canal; fetal gender; prenatal testing; single-cell analysis; trophoblast.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Academic Medical Centers
  • Biomarkers / metabolism
  • Cell Lineage
  • Cells, Cultured
  • Cervix Uteri / metabolism*
  • Chorionic Gonadotropin, beta Subunit, Human / metabolism
  • Chromosomes, Human, X
  • Chromosomes, Human, Y
  • DNA Fragmentation
  • Double-Blind Method
  • Female
  • Genetic Testing
  • Genotype
  • Gestational Age
  • Humans
  • Immunomagnetic Separation*
  • In Situ Hybridization, Fluorescence
  • In Situ Nick-End Labeling
  • Male
  • Phenotype
  • Polymerase Chain Reaction
  • Predictive Value of Tests
  • Pregnancy
  • Prenatal Diagnosis / methods*
  • Prospective Studies
  • Reproducibility of Results
  • Sex Determination Analysis*
  • Trophoblasts / metabolism*

Substances

  • Biomarkers
  • Chorionic Gonadotropin, beta Subunit, Human