Viability and intracellular nitric oxide generation in the umbilical cord blood CD34(+)CD133(–) and CD34(+)CD133(+) cell populations exposed to local anaesthetics

Local anesthetics (LAs) are capable of influencing cell viability in systemic immunity and may also modify metabolism of those present in umbilical cord blood (UCB) following obstetric neuraxial analgesia and anaesthesia. Data regarding UCB immature cells, important for the neonate and critical for putative UCB transplantations, are lacking. LAs are capable of stimulating intracellular nitric oxide (NO) in human neutrophils; no information is available concerning newly perpetuated cells and its potential association with viability. The study aimed at assessing the LAs influence on the cell viability and intracellular NO production by UCB CD34(+)CD133(–) and CD34(+) CD133(+) cell populations. Mononuclear cells separated from UCB samples (n = 19) were incubated with bupivacaine (0.0005, 0.005, 1 mM), lidocaine (0.002, 0.02, 4 mM), and ropivacaine (0.0007, 0.007, 1.4 mM) for 4 h. Flow cytometry was applied for the assessment of cell viability and intracellular NO generation in CD34(+)CD133(–) and CD34(+)CD133(+) cell populations using annexinV/7-AAD and DAF-2DA stainings, respectively. CD34(+)CD133(+) cells showed less pronounced late apoptosis and necrosis as compared to CD34(+)CD133-population. Intracellular NO generation was comparable between both cell populations studied. LAs neither influenced cell viability nor changed NO production in either population. LAs do not interfere with viability and intracellular NO generation in the UCB CD34(+)CD133(–) and CD34(+)CD133(+) cell populations.