Significance of cord blood nucleated red blood cell count in pregnancies complicated by preeclampsia as a marker for intra uterine growth restriction

Background: Preeclampsia is a pregnancy-specific syndrome of reduced organ perfusion secondary to vasospasm and endothelial activation which is typically characterized by hypertension, proteinuria, edema and fetal compromise which is a leading cause of intra uterine growth restriction (IUGR).  Elevated nucleated red blood cell (NRBC) count is introduced as a potential marker of intra-uterine growth restriction (IUGR) hence determination of NRBC counts is essentially helpful in predicting short term neurodevelopment outcome. Objective of the study Elevated nucleated red blood cell (NRBC) count is introduced as a potential marker of intra-uterine growth restriction (IUGR) in term babies born to preeclamptic mothersMethods: A cross sectional study conducted in a tertiary care hospital to evaluate the significance of cord blood NRBC count in term neonates born with pregnancy complicated by preeclampsia. It included 60 healthy mothers (control group) and 60 mothers with PIH. Collected data was analysed with SPSS software.Results: The nucleated red blood cell in cord blood of newborns in preeclampsia group was significantly higher than in the control group (p value 0.013). IUGR is significantly higher in PIH group (p value 0.008).Conclusions: From the observed data it is concluded that IUGR is an important cause of perinatal morbidity and mortality. The commonest maternal cause for IUGR was pregnancy induced hypertension. The other contributing factors were anemia, lack of awareness (unregistered, unbooked cases) among mothers, poor maternal nutrition and poor weight gain during pregnancy.  Infants of preeclamptic women have higher nucleated red blood cell count at birth than control which means that preeclampsia may produce an erythropoietic response in the fetus. The positive correlation between cord nucleated red blood cell counts in preeclamptic patients and control group indicates that may be the hypoperfused placenta plays a role in this correlation.

Comparison of anti-gamma hemoglobin antibody and CD71 antibodies in isolation of fetal nucleated erythrocytes from maternal blood / 대한산부인과학회지

OBJECTIVE: In an attempt to further maximize the potential of genetic analysis from fetal cells isolation, fetal nucleated red blood cell (FNRBC) recovery with direct anti-gamma hemoglobin staining after density gradient and depletion was compared with three different whole blood magnetic separations (1-step and 2-step ferrofluid, 2-step Dynal beads). METHODS: In model systems such as quantitatively defined spikes of fetal into adult blood, as well as blood samples after surgical termination procedures, fetal cell yield and purity through the results of fluorescence in situ hybridization (FISH), quantitative real time polymerase chain reaction (PCR), and fluorescence-activated cell sorting (FACS) were calculated. RESULTS: The yield of total number of cells with a XY signal after FISH was the highest on direct anti-gamma hemoglobin staining. After normalizing the results of each experiment to the corresponding result from anti-gamma hemoglobin staining (1), ratio is 0.42 in 1-step ferrofluid, 0.33 in 2-step ferrofluid, and 0.76 in 2-step dynal beads. The fetal cell purity is clearly better in direct anti-gamma hemoglobin staining than those of the magnetic separations from whole blood. The median ratio is 56.3% in anti-gamma hemoglobin staining, 7.7% in 1-step ferrofluid, 6.5% in 2-step ferrofluid, and 31.4% in 2-step dynal beads. CONCLUSION: This study shows that the direct anti-gamma staining is the best fetal cell recovery system and it is very useful to isolate fetal nucleated red blood cells as a non-invasive genetic source.

Evaluation of NeonatalImmature Reticulocyte Fraction in Small-for-gestational Age Infant in Umbilical Cord Blood / 대한산부인과학회잡지

OBJECTIVE: To evaluate the immature reticulocyte fraction (IRF) by automated calculator that have played a role of predicting marker for hematopoiesis induced by perinatal asphyxia. METHODS: Hospital charts of 40 neonates with diagnosis of small for gestational age (SGA: birth weight <10 percentile) and 47 babies of appropriate for gestational age (AGA) who were delivered vaginally between May 8, 2000 and March 2, 2002 at department of Obstetric and Gynecology, Sanggye Paik Hospital, Inje university. About 5 cc in cord blood was collected immediately after delivery, the calculation of nucleated RBC (N-RBC)/100WBC by macroscopic exam and the evaluation of reticulocyte maturation by automated reticulocyte calculator (Sysmex SE 9000, TOA Medical Electronics Co., Ltd, Kobe, Japan) were performed, and made a comparative study of apgar score. RESULTS: The comparison between AGA and SGA infants, the mean values of cord blood sampling parameter N-RBC (3.53 +/- 4.89 vs 7.92 +/- 10.06, P=0.016), middle fluorescence ratio reticulocyte (MFR) (0.21 +/- 0.02 vs 0.23 +/- 0.03, P<0.001), high fluorescence ratio reticulocyte (HFR) (0.08 +/- 0.03 vs 0.12 +/- 0.03, P<0.001), IRF (0.28 +/- 0.03 vs 0.35 +/- 0.05, P<0.001) were obtained. All of the parameters were significantly different between SGA infants and normal infants and IRF was more valuable marker than N-RBC in evaluation of hematopoietic activity in SGA infants (multiple logistic regression analysis: P value 0.282 vs <0.001). CONCLUSION: Immature reticulocyte fraction by automated calculator is more accurate and confident marker better than N-RBC/100WBC by manual countingin evaluation of the fetal hypoxic induced hematopoiesis.

Obstetrical Outcomes of Increased Nucleated RBC in Maternal Blood during Early Pregnancy / 대한산부인과학회잡지

OBJECTIVE: During early pregnancy, CD71 and glycophorin A positive cells in peripheral blood of pregnant women were studied, to assess the relationship between increased numbers of nucleated RBC (NRBC) in maternal blood and pregnant outcomes. METHODS: Peripheral venous blood samples were obtained from 47 primigravidas of 14~16 weeks gestation. Triple screening tests were routinely performed. Blood samples were incubated with monoclonal anti-CD71 and monoclonal anti-glycophorin antibodies, and analyzed by flow cytometry using FACSort (Becton Dickinson, USA) for checking the NRBC count. RESULTS: A total of 47 pregnant women were enrolled at 14-16 weeks gestation; one pregnancy had anemia and was excluded from the test, the outcome was unknown for 2 other pregnancies, and twelve pregnancies had 1-4% of NRBC in the maternal blood. In the remaining 32 pregnant women, grouped according to their percentage of NRBC, the group with more than 4% of NRBC was termed the study group, and less than 1% of NRBC was termed the control group.The results were as follows: 1) The study group showed lower fetal birth weight than the control group, which was statistically negatively significant (y=-62.219x + 3,401.6, R2=0.2146, p0.05).3) There were two complications in the study group: one was a preterm delivery at 35 weeks of gestational age with birth weight of 2,300 gm and the other was a case of pregnancy-induced hypertension. CONCLUSION: It can be concluded that increased NRBC count in maternal blood during the early second trimester has a significant correlation with fetal birth weight but can't predict high risk pregnancies such as preeclampsia, preterm labor or intrauterine fetal growth restriction.In order to obtain a higher predictive value, further studies with more participants and with high risk pregnancies of known risk factors are needed.

The Correlation between CD34+ Cell and Nucleated RBC Counts in Human Cord Blood

PURPOSE: Human umbilical cord blood as a potential source of hematopoietic stem cell for stem cell transplantation in children has recently been advocated. Clinical application of cord blood transplantation, however, requires adequate blood volume and number of stem cells. Currently, the number of stem cells in the cord blood is usually measured by flowcytometry, which requires strict quality control and high costs. Here, we postulate that the number of nucleated red blood cells(NRBC) which are relatively immature erythroid cells may correlate well with that of CD34-expressing(CD34+) cells which are considered lymphohematopoietic precursors. If so, CD34+ cell-rich cord blood can be selected and such cells enumerated by a simple and cost-effective blood smears. METHODS: Correlation between CD34+ cell and nucleated RBC in human cord blood were checked. Sixty cord blood specimens(30 specimens for group A with NRBC>500/mul and 30 specimens for group B with NRBC500/ul; 55+/-61(SD))(p<0.005). 3.Regression relationship among CD34+ cell, WBC, and NRBC counts was as following. ln(CD34+ cell) = -12.21 + 1.46xln(WBC)+ 0.25xln(NRBC) gamma2=30.07 p=0.0001 4.There was a significant correlation between CD34+ cell counts and NRBC counts(gamma=0.4334, p<0.005) CONCLUSIONS: Our results suggest that there is a significant correlation between CD34+ cell counts and NRBC counts, and that CD34+ cell-rich cord blood specimens can be selected by a simple NRBC counting.