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Brain research bulletin

Organotypic slices culture model for cerebellar ataxia: potential use to study Purkinje cell induction from neural stem cells.


PMID 21145378

Abstract

Cerebellar ataxia is a high profile cerebella disorder currently without viable treatment. In order to establish an in vitro model of cerebellar ataxia for evaluating the potential of neural stem cells (NSC) to repair damage caused by cerebella disorders, organotypic slices of cerebellum were cultured using Stoppini method. Penitrem A (4 mg/kg) was administered intraperitoneally (IP) to postnatal day 6 neonatal Wistar rats to produce Purkinje cell deficient slices suitable to host transplanted NSCs. The survival and differentiation of lipophilic fluorochrome chloromethylbenzamido dialkylcarbocyanine (CM-DiI) labeled cerebellar NSCs isolated from E14 rat brains were investigated after addition to the slices. Analysis indicated that cerebellar cytoarchitecture was well preserved and Purkinje cells survived for over three weeks with fewer Purkinje cells in slices cultured from Penitrem A treated neonates compared to controls. The transplanted NSCs remained viable when cultured within the cerebellar slices and although the majority remained undifferentiated. Some were DiI/2',3'-cyclic nucleotide 3' phosphodiesterase (CNPase) double positive but no DiI/calbindin double labeled cells were found. Our results show that this model could be used to assess the potential for NSC repair of the damaged cerebellum and dissect the possible underlying mechanisms by observing the interaction between NSCs and Purkinje cells in an easily accessible 3-D environment in vitro.

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