Retrieval of germinal zone neural stem cells from the cerebrospinal fluid of premature infants with intraventricular hemorrhage

Abstract Intraventricular hemorrhage is a common cause of morbidity and mortality in premature infants. The rupture of the germinal zone into the ventricles entails loss of neural stem cells and disturbs the normal cytoarchitecture of the region, compromising late neurogliogenesis. Here we demonstrate that neural stem cells can be easily and robustly isolated from the hemorrhagic cerebrospinal fluid obtained during therapeutic neuroendoscopic lavage in preterm infants with severe intraventricular hemorrhage. Our analyses demonstrate that these neural stem cells, although similar to human fetal cell lines, display distinctive hallmarks related to their regional and developmental origin in the germinal zone of the ventral forebrain, the ganglionic eminences that give rise to interneurons and oligodendrocytes. These cells can be expanded, cryopreserved, and differentiated in vitro and in vivo in the brain of nude mice and show no sign of tumoral transformation 6 months after transplantation. This novel class of neural stem cells poses no ethical concerns, as the fluid is usually discarded, and could be useful for the development of an autologous therapy for preterm infants, aiming to restore late neurogliogenesis and attenuate neurocognitive deficits. Furthermore, these cells represent a valuable tool for the study of the final stages of human brain development and germinal zone biology. Significance statement Intraventricular hemorrhage (IVH), occurring in 15% to 40% of preterm births, is frequently associated with long-term neurological deficits. The rupture of the proliferative germinal zone in IVH disturbs late neuronal, ependymal, and glio-genesis. Using a minimally invasive neuroendoscopic procedure, neural stem cells can be retrieved from the cerebrospinal fluid, which can be expanded, cryopreserved, and differentiated in vitro and in vivo, and are not tumorigenic. These cells display distinct hallmarks related to their origin in the germinal zone of the ventral forebrain and could be useful for the development of an autologous cell therapy aiming to attenuate neurocognitive sequelae.