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New technique using galactose-specific lectin for isolation of fetal cells from maternal blood
Authors:Michihiro Kitagawa  Kentaro Sugiura  Hiroko Omi  Yoshiaki Akiyama  Kiyoko Kanayama  Masaru Shinya  Tadao Tanaka  Hirobumi Yura  Haruhiko Sago
Institution:1. Department of Obstetrics and Gynecology, National Okura Hospital, Tokyo, Japan;2. Department of Obstetrics and Gynecology, Jikei University School of Medicine, Tokyo, Japan;3. Natech Inc., Tokyo, Japan
Abstract:To isolate fetal cells from maternal blood, we developed a new method based on galactose-bearing conjugation. Nucleated red blood cells (NRBCs), which highly express galactose on their surface, were selectively attached to a substrate coated with a galactose-containing polymer via soybean agglutinin (SBA), a galactose-specific lectin. Cord blood samples were used to evaluate enrichment efficacy of NRBCs by this method. Blood samples were obtained from 131 pregnant women between 6 and 27 gestational weeks. After preliminary condensation of fetal cells by Ficoll gradient centrifugation, NRBCs were enriched using galactose-positive selection by adjusting SBA concentration. We isolated one to severalhundred NRBCs (mean±SD, 7.8±8.5) in 2.3 ml of peripheral blood samples from 96% of pregnantwomen. The isolated NRBCs were analyzed by a Y-chromosome FISH probe in eight cases carrying male fetuses. Y-signals were detected in all eight cases and more than half of the NRBCs wereoffetal origin. The study demonstrates that our new method using galactose-specific lectin provides effective enrichment of fetal NRBCs allowing non-invasive prenatal diagnosis. Copyright © 2002 John Wiley & Sons, Ltd.
Keywords:galactose  SBA (soybean agglutinin)  NRBC (nucleated red blood cell)  FCS (fetal cell separation)
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