New technique using galactose-specific lectin for isolation of fetal cells from maternal blood.
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 several hundred NRBCs (mean+/-SD, 7.8+/-8.5) in 2.3 ml of peripheral blood samples from 96% of pregnant women. 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 were off fetal origin. The study demonstrates that our new method using galactose-specific lectin provides effective enrichment of fetal NRBCs allowing non-invasive prenatal diagnosis.
Kitagawa
Michihiro
M
Department of Obstetrics and Gynecology, National Okura Hospital, Tokyo, Japan. kitagawa@sub-tky.go.jp
Sugiura
Kentaro
K
Omi
Hiroko
H
Akiyama
Yoshiaki
Y
Kanayama
Kiyoko
K
Shinya
Masaru
M
Tanaka
Tadao
T
Yura
Hirobumi
H
Sago
Haruhiko
H
D002469:Cell Separation / Q000379:methods*; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D005690:Galactose / Q000378:metabolism*; D005865:Gestational Age; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D037102:Lectins; D008297:Male; D037121:Plant Lectins; D011247:Pregnancy; D030262:Soybean Proteins; D014998:Y Chromosome; D010835:Phytohemagglutinins
D005690:Galactose; D037102:Lectins; D037121:Plant Lectins; D030262:Soybean Proteins; C028824:soybean lectin; D010835:Phytohemagglutinins