Mammalian neurogenic sites are good models for physiological neural cell renewal in the perspective of brain repair. Yet, investigating their stem cell niches is not easy since they are small areas deeply hidden in the brain hemispheres. Organotypic slices could be a useful tool since they substantially retain the three-dimensional tissue organization. The postnatal forebrain subventricular zone (SVZ), as a dynamic structure endowed with proliferation and migration, might undergo striking cellular changes in culture. Literature concerning this ex vivo approach applied to SVZ neurogenic activity and response to damage is scarce and heterogeneous, not considering the fine cellular composition of the slice and not taking into account the modifications occurring as a consequence of the culture conditions. Our aim was to describe in detail what happens in the SVZ when establishing an ex vivo model. We addressed the changes occurring in five day-old, postnatal mice forebrain organotypic slices cultured for several days in vitro, by using confocal and ultrastructural analyses. We found that during the first two days in vitro the slices undergo progressive structural disaggregation accompanied by remarkable increase in cell proliferation and death with respect to basal levels. In addition, these facts occur in parallel with strong activation of astrocytic cells and microglia. Our results highlight technical limits in the use of forebrain organotypic slices for studying the activity of SVZ neurogenic niche, indicating that they can be reliable for a very short time (1-2 days) and could be misleading when addressing lesion-induced responses.
Culturing conditions remarkably affect viability and organization of mouse subventricular zone in ex-vivo cultured forebrain slices
ARMENTANO, MARIA;CROCIARA, PAOLA;BONFANTI, Luca
2011-01-01
Abstract
Mammalian neurogenic sites are good models for physiological neural cell renewal in the perspective of brain repair. Yet, investigating their stem cell niches is not easy since they are small areas deeply hidden in the brain hemispheres. Organotypic slices could be a useful tool since they substantially retain the three-dimensional tissue organization. The postnatal forebrain subventricular zone (SVZ), as a dynamic structure endowed with proliferation and migration, might undergo striking cellular changes in culture. Literature concerning this ex vivo approach applied to SVZ neurogenic activity and response to damage is scarce and heterogeneous, not considering the fine cellular composition of the slice and not taking into account the modifications occurring as a consequence of the culture conditions. Our aim was to describe in detail what happens in the SVZ when establishing an ex vivo model. We addressed the changes occurring in five day-old, postnatal mice forebrain organotypic slices cultured for several days in vitro, by using confocal and ultrastructural analyses. We found that during the first two days in vitro the slices undergo progressive structural disaggregation accompanied by remarkable increase in cell proliferation and death with respect to basal levels. In addition, these facts occur in parallel with strong activation of astrocytic cells and microglia. Our results highlight technical limits in the use of forebrain organotypic slices for studying the activity of SVZ neurogenic niche, indicating that they can be reliable for a very short time (1-2 days) and could be misleading when addressing lesion-induced responses.
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