Noel feto-specific mRNA species suitable for identification of fetal cells from the maternal circulation
Noel feto-specific mRNA species suitable for identification of fetal cells from the maternal circulation
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DOI：
10.1002/pd.141
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发表时间：
2001-10-01
影响因子：
3
通讯作者：
Christiansen, M
中科院分区：
文献类型：
--
作者：
Boye, K;Hougaard, DM;Christiansen, M
胎儿特异性标记物是从母血中分离的胎儿细胞进行遗传学诊断所必需的。差异显示逆转录-聚合酶链反应（DDRT-PCR）被用来确定优先表达的mRNA种类滋养层富集的原代细胞培养相比，女性外周血单核细胞（FPMBC）。我们获得了15个不同的cDNA克隆主要在胎盘细胞中表达，使用商业试剂盒与寡聚（dT）锚引物和任意上游引物的总RNA的差异显示分析。反向北方斑点杂交分析和半定量RT-PCR证实了不同的表达水平。7种mRNA仅在胎盘中表达：β-珠蛋白（一种著名的胎儿成红细胞标志物）、早期胎盘胰岛素样肽（EPIL）、YAP 65和骨桥蛋白的mRNA高水平表达，而PP 14、JM 27蛋白和一种未鉴定的表达序列标签（EST）的mRNA中度表达。另外8种mRNA在FPMBC中低表达，在妊娠早期胎盘中高表达。一些mRNA在滋养层细胞系中表达，例如YAP 65，而其他mRNA则不表达，例如EPIL。新的mRNA标记物可用于构建DNA/RNA探针，以通过杂交技术鉴定母体循环中的胎儿细胞，或鉴定胎儿细胞中选择性表达的蛋白质并进行免疫细胞化学检测。版权所有（C）2001约翰威利父子有限公司
Feto-specific markers are necessary for genetic diagnostics on fetal cells isolated froth maternal blood. Differential display reverse transcription-polymerise 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. Copyright (C) 2001 John Wiley & Sons, Ltd.