Purwosunu Yuditiya, Sekizawa Akihiko, Farina Antonio, Okai Takashi, Takabayashi Haruo, Wen Peng, Yura Hirofumi, Kitagawa Michihiro
Department of Obstetrics and Gynecology, Showa University School of Medicine, Tokyo, Japan.
Prenat Diagn. 2006 Jun;26(6):545-7. doi: 10.1002/pd.1456.
To assess the efficiency and reliability of the separation of fetal nucleated red blood cells (NRBCs) using the galactose-specific lectin method, we counted the number of NRBCs in the blood of pregnant women at various gestational ages, as well as after amniocentesis and termination.
Peripheral blood samples were obtained from (1) 22 singleton pregnant women (between 9 and 34 weeks of gestation) and from 23 women who underwent termination (between 6 and 19 weeks of gestation). To determine whether amniocentesis influences numbers of NRBCs, five samples were obtained (2) before and after the procedure. NRBC enrichment was initially performed using density gradients and subsequently using galactose-specific lectin. The cells were then stained with May-Gruenwald Giemsa (MGG) and counted under a light microscope.
NRBCs were found in all samples, ranging from 1 to 82 (median = 12.5 cells/sample). The multiples of the median (MoM) conversion of the number of cells revealed a raise of 1.66-fold (0.12-6.64) in post-termination samples compared with the control value of 1.00 MoM (0.11-6.92; p = 0.036). The postamniocentesis increase was, instead, 1.11-fold (0.17-4.02), which did not reach statistical significance.
All blood samples tested contained NRBCs. Samples obtained after termination yielded more cells than those obtained from women whose pregnancies were going on normally. The number of NRBCs in post-termination samples after MoM conversion differed significantly from those in controls. Although separation of NRBCs was not feasible due to extremely low numbers, our results indicated that NRBCs are detectable in all blood samples from normal pregnant women.
为评估使用半乳糖特异性凝集素方法分离胎儿有核红细胞（NRBCs）的效率和可靠性，我们对不同孕周孕妇以及羊膜穿刺术后和终止妊娠后的血液中的NRBCs数量进行了计数。
采集了外周血样本，其中包括（1）22名单胎孕妇（妊娠9至34周）和23名接受终止妊娠的女性（妊娠6至19周）。为确定羊膜穿刺术是否会影响NRBCs数量，在该操作前后采集了5份样本。首先使用密度梯度法进行NRBC富集，随后使用半乳糖特异性凝集素。然后将细胞用May-Gruenwald Giemsa（MGG）染色，并在光学显微镜下计数。
在所有样本中均发现了NRBCs，数量范围为1至82个（中位数=12.5个细胞/样本）。细胞数量的中位数倍数（MoM）转换显示，终止妊娠后的样本与对照值1.00 MoM（0.11至6.92；p=0.036）相比增加了1.66倍（0.12至6.64）。相反，羊膜穿刺术后的增加为1.11倍（0.17至4.02），未达到统计学显著性。
所有测试的血液样本中均含有NRBCs。终止妊娠后获得的样本比正常妊娠女性获得的样本产生的细胞更多。MoM转换后终止妊娠样本中的NRBCs数量与对照组有显著差异。尽管由于数量极低，NRBCs的分离不可行，但我们的结果表明，正常孕妇的所有血液样本中均可检测到NRBCs。
