Comparision in the yield of fetal nucleated red blood cell between the first-and second-trimester using double density gradient centrifugation.
10.5468/kjog.2010.53.2.127
- Author:
Yeon Jin JEON
1
;
Kwon Hoon KWON
;
Jong Won KIM
;
Myung Geol PANG
;
Sung Chul JUNG
;
Young Ju KIM
Author Information
1. Department of Biochemistry, Ewha Womans University, School of Medicine, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Fetal nucleated red blood cell;
Fetal cell isolation;
Magnetic activated cell sorting;
Double density gradient system;
Prenatal diagnosis
- MeSH:
Aneuploidy;
Centrifugation, Density Gradient;
Erythrocytes;
Female;
Fluorescence;
Humans;
In Situ Hybridization;
Magnetics;
Magnets;
Osmolar Concentration;
Povidone;
Pregnancy;
Pregnancy Trimester, First;
Pregnancy Trimester, Second;
Pregnant Women;
Prenatal Diagnosis;
Silicon Dioxide;
Y Chromosome
- From:Korean Journal of Obstetrics and Gynecology
2010;53(2):127-136
- CountryRepublic of Korea
- Language:English
-
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.