Objectives Intraventricular Haemorrhage (IVH) impacts on ependyma and neural stem cells (NSC) within the subventricular zone.1 Developing therapeutic interventions to curtail this impact is a key research goal. Organotypic models from the wall of the lateral ventricle (LV) of mice2 and humans have the potential to offer unique insight into the dynamic impact of IVH.
Design Basic science research using whole mount preparations (WMP) from the wall of the LV from mice and humans (SOLVe trial: IRAS ID 247936).
Subjects Transgenic mouse line: Fucci:hGFAPCreER:TomatoTD. Human samples from children undergoing surgery for medically intractable epilepsy.
Methods Using a published protocol2 WMP of the LV were prepared. Two samples of human CSF were acquired: a ‘haemorrhagic’ sample from a neonate with IVH and a ‘clean’ sample from a newborn undergoing repair of myelomeningocele. 72 hours time-lapse confocal microscopy was undertaken from multiple wells.
Results Colocalisation of GFP (Fucci+ve) in hGFAP +ve (RFP +ve) cells revealed dividing NSC within the wall of the LV. Quantification of fucci +ve cells revealed ‘haemorrhagic’ CSF caused a significant reduction in proliferation whilst ‘clean’ CSF caused a significant increase in early proliferation.
Conclusions Organotypic slice preparation from mice and human lateral ventricle represents a novel approach to investigating the impact of IVH on the wall of the lateral ventricle.
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