Fetal nucleated erythrocyte recovery: fluorescence activated cell sorting-based positive selection using anti-gamma globin versus magnetic activated cell sorting using anti-CD45 depletion and anti-gamma globin positive selection.
BACKGROUND
Fluorescence activated cell sorting (FACS)-based anti-gamma (gamma) positive selection and magnetic activated cell sorting (MACS)-based anti-CD45 depletion followed by anti-gamma positive staining have been two of the most frequently used methods to isolate fetal cells from maternal blood. To date, there has been no direct comparison of fetal cell recovery by these two methods. This study was designed to address this issue.


METHODS
Fluorescence in situ hybridization (FISH) was performed on nucleated anti-gamma positive cells using X and Y probes. Twenty-four maternal blood samples were obtained immediately after elective termination of pregnancy to ensure a detectable number of fetal cells.


RESULTS
The yield and purity of fetal nucleated erythrocytes (FNRBCs) was statistically higher in FACS sorted samples (P < 0.01). The specificity of staining for FNRBCs was statistically higher in MACS sorted samples (P < 0.01).


CONCLUSIONS
The data from this study demonstrate that both techniques have benefits and limitations. FACS has the advantage of having higher yield, higher purity, higher FISH efficiency and ease in microscope analysis, and MACS has the advantage of having higher specificity and less cell loss during FISH.
Wang
J Y
JY
Division of Genetics, Department of Pediatrics, New England Medical Center, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
Zhen
D K
DK
Falco
V M
VM
Farina
A
A
Zheng
Y L
YL
Delli-Bovi
L C
LC
Bianchi
D W
DW
D015703:Antigens, CD / Q000032:analysis; D002467:Cell Nucleus / Q000648:ultrastructure; D002469:Cell Separation / Q000379:methods; D004622:Embryo, Mammalian; D004912:Erythrocytes / Q000166:cytology* / Q000648:ultrastructure; D005260:Female; D005312:Fetal Blood / Q000166:cytology; D005333:Fetus; D005434:Flow Cytometry / Q000379:methods*; D005865:Gestational Age; D005914:Globins / Q000032:analysis*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods; D017404:In Situ Hybridization, Fluorescence; D017493:Leukocyte Common Antigens / Q000032:analysis; D011247:Pregnancy
D015703:Antigens, CD; D005914:Globins; D017493:Leukocyte Common Antigens