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Inconsistency of fetal trophoblast cells in first trimester maternal peripheral blood prevents non-invasive fetal testing using this cell target.

机译:孕早期孕妇外周血中胎儿滋养层细胞的不一致会阻止使用该细胞靶进行无创胎儿检测。

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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.
机译:我们已经研究了妊娠初期的孕妇外周血是否是可鉴定的滋养细胞的可靠来源。从30 ml静脉血中富集细胞,先前显示了多种抗体可以富集滋养细胞和基于已知滋养细胞表面特征的新混合物。测试了三种不同的磁性固相以富集滋养细胞,并研究了阳性和阴性细胞富集策略。通过形态学结合免疫细胞化学将细胞角蛋白与三种IGF-II,PAI-1或hPLH蛋白之一共定位,或通过与50种针对8种不同表达基因的寡核苷酸的混合物进行原位杂交,将细胞鉴定为滋养细胞-HCG,IGF-II,PAI-1,HASH2,hPLH,p57(KIP2),PP5,H-19。尽管这些工具在绒毛膜绒毛细胞/芽苗制备物和组织切片中都能很好地工作,但即使在终止妊娠后5-20分钟内或从维持妊娠的个体中抽取了母体外周血,我们也无法检测和鉴定出母体外周血中的滋养细胞根据我们自己的经验以及文献中的一些报道,滋养细胞似乎不是以母亲外周血为来源进行胎儿筛查的可行候选物。重要的是要注意,尽管滋养层细胞驱逐是一种可重复描述的生物学现象,但它们不提供进行产前遗传学诊断的方法。

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