| Endothelial progenitor cells (EPCs) are precursor cells of endothelial cells with high proliferative potential, which not only participate in embryonic angiogenesis but also play an important role in adult neovascularization. Many researches concerning adult cardiovascular disease are conducted but less research about neonatal aspects. Sugawara et al found that the number of EPCs in maternal peripheral blood increased along with pregnancy, indicating that EPCs may play a vital role in maintaining placentation during pregnancy and angiogenesis. Acosta et al found that EPC both in pregnant women with gestational diabetes and postpartum umbilical cord blood were reduced, suggesting that changes of number and function of EPCs in maternal peripheral blood may reflect some kind of pathological pregnancy state. Baker et al found that the number of EPC in umbilical cord blood of premature infants was greater than full-term and the number of EPC in umbilical cord blood was reduced among newborns with bronchopulmonary dysplasia. Machalinska et al found that the number of EPC in umbilical cord blood increased significantly among premature infants with ROP. The above mentioned suggests that there will be corresponding changes of EPCs among pregnant women or sick newborns. Therefore, we studied the difference of the number of EPCs in umbilical cord blood between premature infants and full-term clarifying that whether it can provide theoretical data for the predication of fetal disease, severity assessment and other aspects.Objective:To investigate the changes of the number of EPCs in umbilical cord blood between premature and cesarean infants providing a theoretical basis for diseases premature and cesarean infants are susceptible to.Methods:From October2012to September2013,13cases of umbilical cord blood of premature infants were collected (all single births,30to36weeks gestational age,6cases of cesarean section,7cases of natural delivery,6males,7females).24cases of umbilical cord blood of full-term infants were collected (all single births,37to43weeks gestational age,13cases of cesarean section,11cases of natural delivery,14males and10females).1ml umbilical cord blood samples were collected at birth within10minutes, placed in heparin tube and preserved in low temperature. These samples were processed within3hours for machine detection. Flow cytometer was used to analyze EPC.Specific steps:(1) Took sample of100μl anticoagulant umbilical cord blood,10μlCD133-PE antibody and10μl VEGFR-2-APC antibody. After vibration, the sample was incubated at room temperature in the dark for20min.(2) lml hemolysin was added. After vibration, the sample was kept in dark at room temperature for5min.(3) After 1500rpm centrifugation5min, the supernatant was discarded.(4) Adding1ml PBS, vibration,1500rpm centrifugation5min, the supernatant was discarded.(5) Adding0.5ml PBS, the sample was analyzed by flow cytometer. Throughout the study, the analysis was performed and recorded by the same technician who did not know the source of specimen information at the flow chamber, Jinhua Central Hospital.500,000cells were collected from the test specimens. The ratio of the number of EPC during circulation to individual nucleus cells represented as experimental data. Both CD133and VEGFR-2double-antibody labeled positive were considered as target EPC. Using flow cytometer to analyze EPC in umbilical cord blood of premature infants and full-term respectively.Results:EPC in umbilical cord blood of premature infants accounted for0.013%±0.0124%of individual nucleus cells. EPC in umbilical cord blood of full-term infants accounted for0.0036%±0.0025%of individual nucleus cells. The difference between the two groups was statistically significant (t=3.614, P=0.001). EPC in umbilical cord blood of full-term cesarean infants accounted for0.0027%±0.0020%of individual nucleus cells. EPC in umbilical cord blood of full-term natural delivery infants accounted for0.0048%±0.0026%of individual nucleus cells. The difference between two groups was statistically significant (t=2.226,P=0.039). EPC in umbilical cord blood of full-term baby boys and baby girls accounted for0.0042%±0.0024%,0.0036%±0.0030%of individual nucleus cells respectively. The difference between two groups was not statistically significant (t=0.590, P=0.477). EPC in umbilical cord blood of premature cesarean and natural delivery infants accounted for0.0126%±0.0159%,0.0134%± 0.0097%of individual nucleus cells respectively. The difference of the two groups was not statistically significant (t=0.104, P=0.920). EPC in umbilical cord blood of premature baby boys and baby girls accounted for0.0134%±0.0157%,0.0126%±0.010%of individual nucleus cells respectively. The difference of the two groups was not statistically significant (t=0.108, P=0.917). |
PDF Full Text Request