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Obstet Gynecol Sci > Volume 53(2); 2010 > Article
Korean Journal of Obstetrics & Gynecology 2010;53(2):127-136.
DOI: https://doi.org/10.5468/kjog.2010.53.2.127    Published online February 1, 2010.
Comparision in the yield of fetal nucleated red blood cell between the first-and second-trimester using double density gradient centrifugation.
Yeon Jin Jeon, Kwon Hoon Kwon, Jong Won Kim, Myung Geol Pang, Sung Chul Jung, Young Ju Kim
1Department of Biochemistry, Ewha Womans University, School of Medicine, Seoul, Korea.
2GenDix Inc., Seoul, Korea.
3Biomedlab Co., Seoul, Korea.
4Department of Animal Science & Technology and BET Research Institute, Chung-Ang University, Gyeonggi-Do, Korea.
5Department of Obstetrics and Gynecology, Ewha Womans University, School of Medicine, Seoul, Korea.
6Medical Research Institute, Ewha Womans University, Seoul, Korea. kkyj@ewha.ac.kr
Abstract
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.
Key Words: Fetal nucleated red blood cell, Fetal cell isolation, Magnetic activated cell sorting, Double density gradient system, Prenatal diagnosis
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