Ketamine alters the neurogenesis of rat cortical neural stem progenitor cells

Objective: High doses or prolonged exposure to ketamine increase neuronal apoptosis in the developing brain, although effects on neural stem progenitor cells remain unexplored. This study investigated dose-and time-dependent responses to ketamine on cell death and neurogenesis in cultured rat fetal cortical neural stem progenitor cells. Design: Laboratory-based study. Setting: University research laboratory. Subject: Sprague-Dawley rats. Interventions: Neural stem progenitor cells were isolated from the cortex of Sprague-Dawley rat fetuses on embryonic day 17. In dose-response experiments, cultured neural stem progenitor cells were exposed to different concentrations of ketamine (0-100 mu M) for 24 hrs. In time-course experiments, neural stem progenitor cells cultures were exposed to 10 mu M ketamine for different durations (0-48 hrs). Measurements and Main Results: Apoptosis and necrosis in neural stem progenitor cells were assessed using activated caspase-3 immunostaining and lactate dehydrogenase assays, respectively. Proliferative changes in neural stem progenitor cells were detected using bromo-deoxyuridine incorporation and Ki67 immunostaining. Neuronal differentiation was assessed using Tuj-1 immunostaining. Cultured neural stem progenitor cells were resistant to apoptosis and necrosis following all concentrations and durations of ketamine exposure tested. Ketamine inhibited proliferation with decreased numbers of bromo-deoxyuridine-positive cells following ketamine exposure to 100 mu M for 24 hrs (p < .005) or 10 mu M for 48 hrs (p < .01), and reduced numbers of Ki67-positive cells following exposure to ketamine concentration > 10 mu M for 24 hrs (p < .001) or at 10 mu M for 48 hrs (p < .01). Ketamine enhanced neuronal differentiation, with all ketamine concentrations increasing Tuj-1-positive neurons (p < .001) after 24-hrs of exposure. This also occurred with all exposures to 10 mu M ketamine for > 8 hrs (p < .001). Conclusions: Clinically relevant concentrations of ketamine do not induce cell death in neural stem progenitor cells via apoptosis or necrosis. Ketamine alters the proliferation and increases the neuronal differentiation of neural stem progenitor cells isolated from the rat neocortex. These studies imply that ketamine exposure during fetal or neonatal life may alter neurogenesis and subsequent brain development. (Crit Care Med 2012; 40:2407-2416)