Durrant L, McDowall K, Holmes R, Liu D
Department of Clinical Oncology, University of Nottingham, UK.
Br J Obstet Gynaecol. 1996 Mar;103(3):219-22. doi: 10.1111/j.1471-0528.1996.tb09709.x.
To isolate fetal trophoblasts and nucleated red blood cells from the peripheral blood of pregnant women.
Trophoblasts were isolated from whole blood of women in the first trimester of pregnancy by a specific monoclonal antibody, 340. Nucleated red blood cells were isolated by separating whole blood on a triple gradient, staining with ferromagnetic particles coated with an antitransferrin monoclonal antibody and separated on a mini magnetic activated cell sorting (MACS) column. Sorted cells were sexed using a nested polymerase chain reaction for a specific sequence on the Y chromosome and the sex was confirmed by karyotyping of chorionic villus samples.
Patients between 10 and 14 weeks of pregnancy who were undergoing elective chorionic villus sampling for the detection of fetal aneuploidies.
Fetal sex determined by polymerase chain reaction on fetal cells sorted from maternal blood.
When both trophoblasts and nucleated red blood cells were sorted, fetal sex was correctly predicted in 12/13 cases (92%), which included correct diagnosis of five of six male pregnancies. More importantly the two techniques were complementary, with only one male pregnancy being diagnosed on both trophoblasts and nucleated red blood cells, two being detected only with trophoblasts and two on nucleated red blood cells alone. No false positives (male signal from a female pregnancy) were diagnosed with either trophoblasts or nucleated red blood cells even with the highly sensitive nested polymerase chain reaction technique, which is very prone to contamination. This study also shows that it is possible to isolate both trophoblasts and nucleated red blood cells from the same sample of maternal blood.
Fetal cells can be isolated from maternal blood at around 10 weeks of pregnancy.
从孕妇外周血中分离胎儿滋养层细胞和有核红细胞。
采用特异性单克隆抗体340从妊娠早期妇女的全血中分离滋养层细胞。通过在三重梯度上分离全血，用抗转铁蛋白单克隆抗体包被的铁磁颗粒染色，并在微型磁激活细胞分选（MACS）柱上分离，从而分离出有核红细胞。使用针对Y染色体上特定序列的巢式聚合酶链反应对分选后的细胞进行性别鉴定，并通过绒毛膜绒毛样本的核型分析来确认性别。
妊娠10至14周、因检测胎儿非整倍体而接受选择性绒毛膜绒毛取样的患者。
通过对从母血中分选的胎儿细胞进行聚合酶链反应确定胎儿性别。
当同时分选滋养层细胞和有核红细胞时，13例中有12例（92%）正确预测了胎儿性别，其中包括6例男性妊娠中的5例诊断正确。更重要的是，这两种技术具有互补性，只有1例男性妊娠在滋养层细胞和有核红细胞中均被检测到，2例仅在滋养层细胞中被检测到，2例仅在有核红细胞中被检测到。即使采用极易受污染的高灵敏度巢式聚合酶链反应技术，无论是滋养层细胞还是有核红细胞均未诊断出假阳性（女性妊娠的男性信号）。本研究还表明，有可能从同一份母血样本中分离出滋养层细胞和有核红细胞。
在妊娠约10周时可从母血中分离出胎儿细胞。
