Enrichment of fetal cells from maternal blood by magnetic activated cell sorting (MACS) with fetal cell specific antibodies: one-step versus two-step MACS.
We report here the results of fetal cell enrichment from maternal blood in 58 pregnant women by the use of magnetic activated cell sorting (MACS) with erythroblast-specific and/or maternal cell specific antibodies. Two approaches were compared; one-step MACS to enrich CD71+ (a membrane-bound marker) or GPA+ (another marker, glycophorin A) fetal cells versus two-step MACS to deplete CD14+ maternal cells and subsequently to enrich fetal (CD71+ or GPA+) cells. The existence of fetal cells was ensured by both FISH with Y-specific probes and karyotyping of respective amniotic and/or chorionic vullus cells, the results being applied for comparison of detection rate for XY fetuses between the two MACS procedures. In 24 (38.8%) of the 58 blood samples examined, Y-positive cells were observed by FISH, whereas there were 38 true XY fetuses later confirmed by karyotyping, including two cases of 47,XY,+21. On the other hand, in Y-negative cells by FISH, there were two cases of 47,XX,+18. The average number of cells sorted did not differ among one-step MACS procedures with anti-CD14, anti-CD71 and anti-GPA antibodies. With the latter, 12 (75%) of 16 Y-positive fetuses were detected, while only one (20%) of 5 Y-positive fetuses was detected by two-step MACS with anti-CD14/anti-GPA antibodies. The detection rate significantly varied (p = 0.0024) between the two procedures, although the numbers of cases examined were small. There was no statistical difference (p > 0.05) between one-step and two-step MACS with other combinations of antibodies. These findings indicate that one-step MACS using the anti-GPA antibody is more effective than two step MACS for enrichment of fetal cells from maternal blood.
Department of Obstetrics and Gynecology, Nagoya City University Medical School, Aichi 467-8601, Japan. zhaoxx@med.nagoya-cu.ac.jp
Xaio Xi
XX
Zhao
Yasuhiko
Y
Ozaki
Nobuhiro
N
Suzumori
Tsuyoshi
T
Sato
Kaoru
K
Suzumori
D000328:Adult; D000918:Antibody Specificity; D015703:Antigens, CD / Q000097:blood / Q000235:genetics; D000944:Antigens, Differentiation, B-Lymphocyte / Q000097:blood / Q000235:genetics; D002874:Chromosome Mapping; D002887:Chromosomes, Human, Pair 18; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D005333:Fetus / Q000276:immunology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D008423:Maternal Age; D011247:Pregnancy; D018566:Pregnancy, High-Risk; D011990:Receptors, Transferrin
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C068126:CD71 antigen; D011990:Receptors, Transferrin