title: Inconsistency of fetal trophoblast cells in first trimester maternal peripheral blood prevents non-invasive fetal testing using this cell target.
abstract: We have investigated whether maternal peripheral blood from the first trimester of pregnancy is a reliable source of identifiable trophoblast cells. The cells were enriched from 30 ml of venous blood, with multiple antibodies shown previously to enrich trophoblasts and a new cocktail based on known trophoblast surface features. Three different magnetic solid phases were tested to enrich trophoblasts, and both positive and negative cell enrichment strategies were examined. The cells were identified as trophoblast by morphology coupled with immunocytochemistry to co-localize cytokeratin with one of three IGF-II, PAI-1 or hPLH proteins or by in-situ hybridization with a mixture of 50 oligos directed to eight different expressed genes, alpha-HCG, IGF-II, PAI-1, HASH2, hPLH, p57(KIP2), PP5, H-19. While these tools worked beautifully in chorionic villi cell/sprout preparations and tissue sections, we could not detect and identify any trophoblasts in maternal peripheral blood even if the maternal peripheral blood was drawn 5-20 min following termination of pregnancy or from individuals maintaining the pregnancy. Based on our own experience and that of some reports in the literature, trophoblasts do not appear to be a viable candidate for fetal screening using maternal peripheral blood as the source. It is important to note that while trophoblast deportation is a biological phenomenon that has been described repeatable, they do not provide a means to perform prenatal genetic diagnosis.
authors.affiliation: Chief Technology Office, Roche Diagnostics, Berkeley, CA, USA.
authors.forename: P A
authors.initials: PA
authors.lastname: Schueler
authors.forename: D T
authors.initials: DT
authors.lastname: Yamanishi
authors.forename: J
authors.initials: J
authors.lastname: Pearson
authors.forename: Y
authors.initials: Y
authors.lastname: Lee
authors.forename: X
authors.initials: X
authors.lastname: Wu
authors.forename: S
authors.initials: S
authors.lastname: Hashima
authors.forename: M R
authors.initials: MR
authors.lastname: Madlansacay
authors.forename: C A
authors.initials: CA
authors.lastname: Cain
authors.forename: E J
authors.initials: EJ
authors.lastname: Collarini
authors.forename: L
authors.initials: L
authors.lastname: Foltz
authors.forename: W
authors.initials: W
authors.lastname: Mahoney
mesh_terms: D000906:Antibodies; D051792:Basic Helix-Loop-Helix Proteins; D001773:Blood Cells / Q000166:cytology*; D001943:Breast Neoplasms; D002469:Cell Separation; D004268:DNA-Binding Proteins / Q000235:genetics; D066246:ErbB Receptors / Q000276:immunology; D005260:Female; D005656:Fungal Proteins / Q000235:genetics; D015292:Glycoprotein Hormones, alpha Subunit / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007335:Insulin-Like Growth Factor II / Q000032:analysis / Q000235:genetics; D007633:Keratins / Q000032:analysis; D007962:Leukocytes; D008280:Magnetics; D008869:Microtubule-Associated Proteins / Q000235:genetics; D020409:Molecular Motor Proteins; D017395:Plasminogen Activator Inhibitor 1 / Q000032:analysis / Q000235:genetics; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D012333:RNA, Messenger / Q000032:analysis; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D014327:Trophoblasts / Q000737:chemistry / Q000166:cytology*; D014407:Tumor Cells, Cultured
chemical_list: D000906:Antibodies; D051792:Basic Helix-Loop-Helix Proteins; D004268:DNA-Binding Proteins; D066246:ErbB Receptors; D005656:Fungal Proteins; D015292:Glycoprotein Hormones, alpha Subunit; D007335:Insulin-Like Growth Factor II; D007633:Keratins; D008869:Microtubule-Associated Proteins; D020409:Molecular Motor Proteins; D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C065501:ASCL2 protein, human; C075341:KIP2 protein, S cerevisiae