The application of CD71 and Hoechst33258 to staining method for sorting fetal nucleated red blood cells in the peripheral blood of pregnant women.
OBJECTIVE
To compare mononuclear cells labeled by CD71 single-staining method with CD71 and Hoechst33258(HO258) double-staining one, and sort nucleated red blood cells (NRBCs) in peripheral blood in pregnant women by use of the latter method.


METHODS
Mononuclear cells in umbilical blood and peripheral blood in normal pregnant women and patients with mild or moderate pregnant hypertension syndrome were labeled by erythrocyte specific antibody CD71 and nuclear dye HO258. Furthermore, NRBCs were sorted in combination with the use of flow cytometry.


RESULTS
Labeled by CD71 and Hoechst33258 double-staining method, the positive labeling rate of mononuclear cells in umbilical blood and peripheral blood in normal pregnant women and patients with pregnant hypertension syndrome was remarkably lower than that labeled by CD71 single-staining method (t-test, P<0.01). Meanwhile, CD71 and Hoechst33258 double-staining method was successfully used in sorting NRBCs in peripheral blood in pregnant women by flow cytometry.


CONCLUSIONS
CD71 and HO258 double-staining method could exclude mature erythrocytes, thrombocytes and reticulocytes and this improve NRBCs' specific label and sorting purity.
Tao
D
D
Molecular Medicine Center, Tongji Hospital, Tongji Medical University, Wuhan, Hubei, 430030 P.R.China. dedingtao@hotmail.com
Shen
Y
Y
Feng
X
X
Chen
H
H
D015703:Antigens, CD / Q000276:immunology*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology*; D006690:Bisbenzimidazole / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D011247:Pregnancy / Q000097:blood*; D011990:Receptors, Transferrin; D013194:Staining and Labeling
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C068126:CD71 antigen; D005456:Fluorescent Dyes; D011990:Receptors, Transferrin; D006690:Bisbenzimidazole