A high yield of fetal nucleated red blood cells isolated using optimal osmolality and a double-density gradient system
A high yield of fetal nucleated red blood cells isolated using optimal osmolality and a double-density gradient system
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DOI：
10.1002/pd.1888
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发表时间：
2007-12-01
影响因子：
3
通讯作者：
Pang, M. G.
中科院分区：
文献类型：
--
作者：
Kwon, K. H.;Jeon, Y. J.;Pang, M. G.
目的采用非连续Percoll梯度法分离胎儿有核红细胞(NRBC)，并探讨渗透压对NRBC产率的影响。方法采用不同渗透压的单、双Percoll梯度分离胎儿有核红细胞。采用磁激活细胞分选法对分离的NRBC进行分选，Kleihauer-Betke染色进行形态分化。结果在单密度Percoll色谱柱上，以280mOsm/kg H2O+1.077 g/mLPercoll和520mOsm/kgH2O+1.119 g/mLPercoll分离的NRBC数量最多。用双Percoll密度梯度分离的胎儿NRBC明显多于用双组织不透明密度梯度分离的胎儿NRBC(p=0.043)。对25例NRBC进行FISH分析，正确识别出15例男性、9例女性整倍体胎儿和1例21三体胎儿。结论本文提出的NRBC增殖法与以往的方法相比，所需母血更少，NRBC产量更高。版权所有(C)2007 John Wiley&Sons，Ltd.
Objectives To increase the yield of fetal nucleated red blood cells (NRBCs) from maternal blood using a discontinuous Percoll gradient and to determine the effects of osmolality on NRBC yield.Methods Fetal NRBCs were isolated from combined umbilical cord blood and adult female blood, or from maternal blood using single or double Percoll gradients with different osmolalities. Magnetic activated cell sorting was used to enrich isolated NRBCs, and morphological differentiation was performed with Kleihauer-Betke stain. We also isolated fetal NRBCs from 25 10 mL samples of maternal blood and determined fetal sex by fluorescence in situ hybridization (FISH), using X-Y probes.Results For single-density Percoll columns, the greatest number of NRBCs was isolated using 280 mOsm/kg H2O With 1.077 g/mL Percoll and 520 mOsm/kg H2O With 1.119 g/mL Percoll. Significantly more fetal NRBCs were isolated with double Percoll density gradients than with double-Histopaque gradients (p = 0.043). FISH analysis on NRBC in 25 cases correctly identified 15 male and 9 female euploid fetuses and one Trisomy 21 fetus.Conclusion The NRBC enrichment method we present requires less maternal blood and yields more NRBCs compared to previous methods. Copyright (c) 2007 John Wiley & Sons, Ltd.