APOPTOSIS IN CULTURED SPINAL CORD SLICES OF NEONATAL MOUSE

Abstract

Organotypic spinal cord slices from neonatal mammals could be a powerful model for evaluation of cell survival but also cell death mechanisms. The aim of this study was to establish an in vitro model for investigating cell survival and mechanism involved in cell death in neonatal spinal cord slices. The spinal cord was sliced and incubated into culture medium. The MTT assay was carried out to assess the viability of the slices and fluorescent staining was used to study morphological features of apoptosis, where as nucleosomal DNA fragmentation was detected using agarose gel electrophoresis. The results of the present study demonstrated that the slices could be maintained in culture up to 14 days. Both neurons and glial cells died by apoptosis and application of a general caspase inhibitor neither affected slice survival nor nucleosomal DNA fragmentation after 24 h in culture. In addition, the inhibitor failed to block apoptosis in neurons and glial cells in the cultured slices. Our results suggest that in the cultured slices, apoptosis is the main reason for neuron and glial cell death, which occurs by a caspase-independent mechanism.