Neural stem cell therapy of foetal onset hydrocephalus using the HTx rat as experimental model

Henzi, Roberto; Vío, Karin; Jara, Clara; Johanson, Conrad E.; McAllister, James P.; Rodríguez, Estéban M.; Guerra, Montserrat

doi:10.1007/s00441-020-03182-0

Cited by 17 publications

(12 citation statements)

References 70 publications

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“…The histopathologic consequences of hydrocephalus depend on the age of onset, rate of ventricular enlargement, and the aetiology [2]. However, it has been adjudged not to be only a disorder of CSF dynamics but also a cerebral parenchymal disorder [3] that leads to functional neurological impairments [4,5]. Increase in the CSF pressure together with the expansion of the ventricles stretches and ruptures the ependyma at isolated points, leading, in severe cases, to total destruction of the epithelium [6,7].…”

Section: Introductionmentioning

confidence: 99%

“…Foetal and infantile hydrocephalus occur at a time in development when the neurons are proliferating, and migrating to their final position or forming synapses with other neurons in order to produce functional brain circuits. Human and animal studies have revealed that foetal and perinatal hydrocephalus disrupts the ventricular and sub-ventricular zones causing loss of neural stem cells and the ependyma layer, leading to abnormal neurogenesis [5,8]. Hydrocephalus may also be caused by a malfunction of the ependymal and mesenchymal cells necessary for the signalling pathway and disruption of patterning molecules during early brain development [9].…”

Section: Introductionmentioning

confidence: 99%

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Changes in Neuronal Density of the Sensorimotor Cortex and Neurodevelopmental Behaviour in Neonatal Mice with Kaolin-Induced Hydrocephalus

Femi-Akinlosotu

Shokunbi

2020

Pediatr Neurosurg

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Introduction: Hydrocephalus is a disorder in which the circulation of cerebrospinal fluid is altered in a manner that leads to its accumulation in the ventricles and subarachnoid space. Its impact on the neuronal density and networks in the overlying cerebral cortex in a time-dependent neonatal hydrocephalic process is largely unknown. We hypothesize that hydrocephalus will affect the cytoarchitecture of the cerebral cortical mantle of neonatal hydrocephalic mice, which will in turn modify sensorimotor processing and neurobehaviour. Objective: The purpose of this study is to probe the effect of hydrocephalus on 3 developmental milestones (surface righting reflex, cliff avoidance reflex, and negative geotaxis) and on cortical neuronal densities in neonatal hydrocephalic mice. Methods: Hydrocephalus was induced in 1-day-old mice by intracisternal injection of sterile kaolin suspension. The pups were tested for reflex development and sensorimotor ability using surface righting reflex (PND 5, 7, and 9), cliff avoidance (PND 6), and negative geotaxis (PND 10 and 12) prior to their sacrifice on PND 7, 14, and 21. Neuronal density and cortical thickness in the sensorimotor cortex were evaluated using atlas-based segmentation of the neocortex and boundary definition in 4-μm paraffin-embedded histological sections with hematoxylin and eosin as well as cresyl violet stains. Results: Surface righting and cliff avoidance activities were significantly impaired in hydrocephalic pups but no statistically significant difference was observed in negative geotaxis in both experimental and control pups. The neuronal density of the sensorimotor cortex was significantly higher in hydrocephalic mice than in age-matched controls on PND 14 and 21 (373.20 ± 21.54 × 10−6 μm2 vs. 157.70 ± 21.88 × 10−6 μm2; 230.0 ± 44.1 × 10−6 μm2 vs. 129.60 ± 3.72 × 10−6 μm2, respectively; p < 0.05). This was accompanied by reduction in the cortical thickness (µm) in the hydrocephalic mice on PND 7 (2,409 ± 43.37 vs. 3,752 ± 65.74, p < 0.05), PND 14 (2,035 ± 322.10 vs. 4,273 ± 67.26, p < 0.05), and PND 21 (1,676 ± 33.90 vs. 4,945 ± 81.79, p < 0.05) compared to controls. Conclusion: In this murine model of neonatal hydrocephalus, the quantitative changes in the cortical neuronal population may play a role in the observed changes in neurobehavioural findings.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Changes in Neuronal Density of the Sensorimotor Cortex and Neurodevelopmental Behaviour in Neonatal Mice with Kaolin-Induced Hydrocephalus

Femi-Akinlosotu

Shokunbi

2020

Pediatr Neurosurg

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“…While other authors have shown in vitro congenital hydrocephalus-dependent cytopathology [24,42], this technique is a highly specific method to study the cytological mechanisms involved in the VZ after IVH. In this protocol paper, our aim is to describe our model [32] and show representative results.…”

Section: Discussionmentioning

confidence: 99%

“…An association between IVH, PHH, and neurodevelopmental impairment is well-established, but the mechanisms linking these disorders remain unclear. Recent evidence implicates impairment of cell junction complexes within the ventricular zone (VZ) [ 4 – 10 ] with associated ciliopathy in the etiology of congenital, non-hemorrhagic hydrocephalus [ 11 – 19 ] in both experimental models [ 4 – 6 , 10 , 11 , 20 – 24 ] and humans [ 25 – 28 ]. VZ disruption is also identified as a key feature in IVH in humans [ 29 ].…”

Section: Introductionmentioning

confidence: 99%

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

Castañeyra-Ruiz

McAllister

Morales

et al. 2020

Fluids Barriers CNS

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Background: Severe intraventricular hemorrhage (IVH) is one of the most devastating neurological complications in preterm infants, with the majority suffering long-term neurological morbidity and up to 50% developing post-hemorrhagic hydrocephalus (PHH). Despite the importance of this disease, its cytopathological mechanisms are not well known. An in vitro model of IVH is required to investigate the effects of blood and its components on the developing ventricular zone (VZ) and its stem cell niche. To address this need, we developed a protocol from our accepted in vitro model to mimic the cytopathological conditions of IVH in the preterm infant. Methods: Maturing neuroepithelial cells from the VZ were harvested from the entire lateral ventricles of wild type C57BL/6 mice at 1-4 days of age and expanded in proliferation media for 3-5 days. At confluence, cells were re-plated onto 24-well plates in differentiation media to generate ependymal cells (EC). At approximately 3-5 days, which corresponded to the onset of EC differentiation based on the appearance of multiciliated cells, phosphate-buffered saline for controls or syngeneic whole blood for IVH was added to the EC surface. The cells were examined for the expression of EC markers of differentiation and maturation to qualitatively and quantitatively assess the effect of blood exposure on VZ transition from neuroepithelial cells to EC. Discussion: This protocol will allow investigators to test cytopathological mechanisms contributing to the pathology of IVH with high temporal resolution and query the impact of injury to the maturation of the VZ. This technique recapitulates features of normal maturation of the VZ in vitro, offering the capacity to investigate the developmental features of VZ biogenesis.

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“…The cells are expanded for 3-5 days or until con uence. While other authors have shown in vitro congenital hydrocephalus-dependent cytopathology (24,42), this technique is a highly speci c method to study the cytological mechanisms involved in the VZ after IVH. In this protocol paper, our aim is to describe our model (32) and show representative results.…”

Section: Discussionmentioning

confidence: 99%

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

Castañeyra-Ruiz¹,

McAllister

Morales

et al. 2020

Preprint

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Background: Severe intraventricular hemorrhage (IVH) is one of the most devastating neurological complications in preterm infants, with the majority suffering long-term neurological morbidity and up to 50 percent developing post-hemorrhagic hydrocephalus (PHH). Despite the importance of this disease, its cytopathological mechanisms are not well known. An in vitro model of IVH is required to investigate the effects of blood and its components on the developing ventricular zone (VZ) and its stem cell niche. To address this need, we developed a protocol from our accepted in vitro model to mimic the cytopathological conditions of IVH in the preterm infant. Methods: Maturing neuroepithelial cells from the VZ were harvested from the entire lateral ventricles of wild type C57BL/6 mice at 1-4 days of age and expanded in proliferation media for 3-5 days. At confluence, cells were re-plated onto 24-well plates in differentiation media to generate ependymal cells (EC). At approximately 3-5 days, which corresponded to the onset of EC differentiation based on the appearance of multiciliated cells, phosphate-buffered saline for controls or syngeneic whole blood for IVH was added to the EC surface. The cells were examined for the expression of EC markers of differentiation and maturation to qualitatively and quantitatively assess the effect of blood exposure on VZ transition from neuroepithelial cells to EC. Discussion: This protocol will allow investigators to test cytopathological mechanisms contributing to the pathology of IVH with high temporal resolution and query the impact of injury to the maturation of the VZ. This technique recapitulates features of normal maturation of the VZ in vitro, offering the capacity to investigate the developmental features of VZ biogenesis.

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Neural stem cell therapy of foetal onset hydrocephalus using the HTx rat as experimental model

Cited by 17 publications

References 70 publications

Changes in Neuronal Density of the Sensorimotor Cortex and Neurodevelopmental Behaviour in Neonatal Mice with Kaolin-Induced Hydrocephalus

Changes in Neuronal Density of the Sensorimotor Cortex and Neurodevelopmental Behaviour in Neonatal Mice with Kaolin-Induced Hydrocephalus

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

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

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