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KMID : 0358420100530020127
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Korean Journal of Obstetrics and Gynecology
2010 Volume.53 No. 2 p.127 ~ p.136
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Comparision in the yield of fetal nucleated red blood cell between the first-and second-trimester using double density gradient centrifugation
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Jeon Yeo-Jin
Kwon Kyoung-Hun
Kim Jong-Won
Pang Myung-Geol
Jung Sung-Chul
Kim Young-Ju
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Abstract
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Objective: The aim of our study was to make a practical comparative evaluation of the first and second trimesters in order to determine the period during which a higher yield of fetal nucleated red blood cells (FNRBCs) can be obtained.
Methods: NRBCs were isolated from maternal blood during the first and second trimesters of pregnancy using double Percoll gradients with different osmolarities. Magnetic activated cell sorting was performed with Kleihauer-Betke stain. We isolated fetal NRBCs from 10 mL of samples of maternal blood and determined fetal sex and fetal aneuploidy by fluorescence in situ hybridization (FISH).
Results: The average number of NRBCs was 9.85 in samples obtained during the first trimester and 14.88 in samples obtained during the second trimester (P=0.07). The average number of NRBCs with Y chromosome signals was 5.73 in the first trimester and 8.22 in second trimester (P=0.56). However, the percentage of NRBCs with Y chromosome signals in the first trimester (70.6%) was significantly higher than in the second trimester (59.8%) (P=0.049). We diagnosed the blood samples from 7 pregnant women having fetal aneuploidy using this method and the number of NRBCs was 18.4.
Conclusion: The method using Percoll osmolarity and a double density gradient system may be a very useful method for separation of NRBCs in the first trimester of pregnancy and also in the second trimester.
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KEYWORD
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Fetal nucleated red blood cell, Fetal cell isolation, Magnetic activated cell sorting, Double density gradient system, Prenatal diagnosis
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