Neurospheres from neural stem/neural progenitor cells (NSPCs) of non-hydrocephalic HTx rats produce neurons, astrocytes and multiciliated ependyma: the cerebrospinal fluid of normal and hydrocephalic rats supports such a differentiation

Abstract

Fetal onset hydrocephalus and abnormal neurogenesis are two inseparable phenomena turned on by a cell junction pathology first affecting neural stem/progenitor cells (NSPCs) and later the multiciliated ependyma. The neurological impairment of children born with hydrocephalus is not reverted by derivative surgery. NSPCs and neurosphere (NE) grafting into the cerebrospinal fluid (CSF) of hydrocephalic fetuses thus appears as a promising therapeutic procedure. There is little information about the cell lineages actually forming the NE as they grow throughout their days in vitro (DIV). Furthermore, there is no information on how good a host the CSF is for grafted NE. Here, we use the HTx rat, a model with hereditary hydrocephalus, with the mutation expressed in about 30% of the litter (hyHTx), while the littermates develop normally (nHTx). The investigation was designed (i) to establish the nature of the cells forming 4 and 6-DIV NE grown from NSPCs collected from PN1/nHTx rats and (ii) to study the effects on these NEs of CSF collected from nHTx and hyHTx. Immunofluorescence analyses showed that 90% of cells forming 4-DIV NEs were non-committed multipotential NSPCs, while in 6-DIV NE, 40% of the NSPCs were already committed into neuronal, glial and ependymal lineages. Six-DIV NE further cultured for 3 weeks in the presence of fetal bovine serum, CSF from nHTx or CSF from hyHTx, differentiated into neurons, astrocytes and βIV-tubulin+ multiciliated ependymal cells that were joined together by adherent junctions and displayed synchronized cilia beating. This supports the possibility that ependymal cells are born from subpopulations of NSC with their own time table of differentiation. As a whole, the findings indicate that the CSF is a supportive medium to host NE and that NE grafted into the CSF have the potential to produce neurons, glia and ependyma.

Preterm Intraventricular Hemorrhage in vitro: Modeling the Cytopathology of the Ventricular Zone

“…An increase in multiciliated ependymal cells over time is detected in control conditions (Fig 2 A-B' & F). To con rm this, we quanti ed the percentage of cells expressing EMA on days 5 (37.51 ± 11.38) , 6 (42.93 ± 10.77), and 7 (58.15 ± 10.66) (supplemental Fig 1); these percentages were similar to those previously reported by our group (41).…”

“…In fact, de cits in the formation of cilia cause serious disorders in the central nervous system, including hydrocephalus (reviewed in (36)(37)(38)). Despite the importance of the VZ in brain development, there are few reported methods to differentiate EC from brain neural stem cells (NSC) (29,(39)(40)(41). We have recently reported on the effects of syngeneic blood on the VZ using an in vitro model (42), but the details of our procedures have not been published.…”

Brain ependymocytes in neurogenesis and maintaining integrity of blood-cerebrospinal fluid barrier

“…Кроме того, наблюдается ассиметричное деление эпендимальных клеток, которое характерно для НСК, а среди клеток, которые образуются из делящихся эпендимальных клеток, присутствуют нейроны, астроциты и олигодендроциты [25,45]. При этом в случае аномального нейрогенеза сначала поражаются НСК и клетки-предшественники, а затем эпендимоциты [46]. В эксперименте доказано, что частичное разрушение эпендимоцитов путем интрацеребровентрикулярной инъекции нейраминидазы привело к увеличению количества нейробластов в субвентрикулярной зоне, что доказывает контролирующую роль эпендимоцитов в пролиферации НСК [47].…”

Preterm intraventricular hemorrhage in vitro: modeling the cytopathology of the ventricular zone

“…In fact, deficits in the formation of cilia cause serious disorders in the central nervous system, including hydrocephalus (reviewed in [ 38 – 40 ]). Despite the importance of the VZ in brain development, there are few reported methods to differentiate EC from brain neural stem cells (NSC) [ 30 , 41 – 43 ]. We have recently reported on the effects of syngeneic blood on the VZ using an in vitro model [ 32 ], but the details of our procedures have not been published.…”