Novel feto-specific mRNA species suitable for identification of fetal cells from the maternal circulation.
Feto-specific markers are necessary for genetic diagnostics on fetal cells isolated from maternal blood. Differential display reverse transcription-polymerase chain reaction (DDRT-PCR) was used to identify mRNA species preferentially expressed in trophoblast-enriched primary cell cultures compared to female peripheral mononuclear blood cells (FPMBCs). We obtained 15 different cDNA clones expressed predominantly in placenta cells using a commercial kit with oligo-(dT) anchor primers and arbitrary upstream primers for differential display analysis of total RNA. Reverse Northern dot-blot analysis and semi-quantitative RT-PCR confirmed the different expression levels. Seven mRNA species were exclusively expressed in the placenta: the mRNAs of epsilon-globin, a renowned marker of fetal erythroblasts, early placenta insulin-like peptide (EPIL), Yes-associated protein (YAP65) and osteopontin were expressed at high levels, whereas the mRNAs of PP14, JM27 protein, and an unidentified expressed sequence tag (EST) were moderately expressed. A further eight mRNAs with low expression in FPMBCs and higher expression in first trimester placenta were identified. Some mRNAs were expressed in a trophoblast cell line, e.g. YAP65, whereas others were not, e.g. EPIL. The new mRNA markers may be used to construct DNA/RNA probes to identify fetal cells in maternal circulation by hybridization techniques, or to identify proteins selectively expressed in fetal cells and amenable to immunocytochemical detection.
Boye
K
K
Department of Clinical Biochemistry, Statens Serum Institut, Copenhagen, Denmark.
Hougaard
D M
DM
Ebbesen
P
P
Vuust
J
J
Christiansen
M
M
D000199:Actins / Q000235:genetics; D048868:Adaptor Proteins, Signal Transducing; D000449:Aldehyde Reductase / Q000235:genetics; D015415:Biomarkers / Q000097:blood*; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000235:genetics; D005260:Female; D005312:Fetal Blood / Q000737:chemistry / Q000166:cytology*; D000071837:Fibrillins; D005914:Globins / Q000235:genetics; D006133:Growth Substances; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007963:Leukocytes, Mononuclear; D008840:Microfilament Proteins / Q000235:genetics; D053495:Osteopontin; D010750:Phosphoproteins / Q000235:genetics; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000097:blood*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000235:genetics; D014157:Transcription Factors; D014327:Trophoblasts / Q000166:cytology; D000091102:YAP-Signaling Proteins
D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D015415:Biomarkers; D002352:Carrier Proteins; D006063:Chorionic Gonadotropin; D000071837:Fibrillins; D006133:Growth Substances; C101126:INSL4 protein, human; D036341:Intercellular Signaling Peptides and Proteins; D008840:Microfilament Proteins; D010750:Phosphoproteins; D011257:Pregnancy Proteins; D012333:RNA, Messenger; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; D014157:Transcription Factors; D000091102:YAP-Signaling Proteins; C088374:YAP1 protein, human; D053495:Osteopontin; D005914:Globins; D000449:Aldehyde Reductase