[Levels of peripheral circulating nucleated erythrocytes in pregnant women for noninvasive prenatal diagnosis].
OBJECTIVE
Enrichment of nucleated red blood cells (NRBCs) from maternal blood for non-invasive prenatal diagnosis.


METHODS
Pilot study.


METHODS
2nd Clinic of Paediatrics, University Hospital Motol, Prague, Czech Republic.


METHODS
Mononuclear cells were isolated from 13-28 ml of peripheral maternal blood between 13 and 37 weeks of gestation. Leukocytes from maternal peripheral blood were depleted from mononuclear cells by treatment with anti-CD14 and anti-CD45 microbeads and high-gradient magnetic cell separation (MACS) on VarioMACS. NRBCs were sorted from CD14-/CD45- fraction by positive selection using anti-CD71 microbeads on MiniMACS. All sorting steps were analysed by three-colour cytometric analysis with FACScan flow cytometer.


RESULTS
In 68 out of 78 pregnant woman (87%) NRBCs were found in range 2 x 10(5) - 1.02 x 10(6). NRBC were enriched with an average enrichment rate of 138-fold ranging from 4-526 fold. In our cohort of pregnant woman the number of isolated NRBCs was individual. We identified NRBCs from the 13th week of gestation.


CONCLUSIONS
The aim of the study is to establish and standardise the method of enrichment of NRBCs from maternal blood samples and verify the applicability of this alternative source for non-invasive prenatal diagnosis.
Hromadníková
I
I
II. dĕtská klinika 2. LF UK, FN Motol, Praha.
Sedlácková
L
L
Mrstinová
M
M
Stechová
K
K
Karamanov
S
S
Kofer
J
J
Macek
M
M
D015703:Antigens, CD / Q000032:analysis; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis; D004900:Erythroblasts / Q000276:immunology; D004906:Erythrocyte Count; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D017493:Leukocyte Common Antigens / Q000032:analysis; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D011247:Pregnancy; D011264:Pregnancy Trimesters; D011296:Prenatal Diagnosis; D011990:Receptors, Transferrin
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C068126:CD71 antigen; D018950:Lipopolysaccharide Receptors; D011990:Receptors, Transferrin; D017493:Leukocyte Common Antigens