Injury Response of Resected Human Brain Tissue In Vitro

Abstract

Brain injury affects a significant number of people each year. Organotypic cultures from resected normal neocortical tissue provide unique opportunities to study the cellular and neuropathological consequences of severe injury of adult human brain tissue in vitro. The in vitro injuries caused by resection (interruption of the circulation) and aggravated by the preparation of slices (severed neuronal and glial processes and blood vessels) reflect the reaction of human brain tissue to severe injury. We investigated this process using immunocytochemical markers, reverse transcriptase quantitative polymerase chain reaction and Western blot analysis. Essential features were rapid shrinkage of neurons, loss of neuronal marker expression and proliferation of reactive cells that expressed Nestin and Vimentin. Also, microglia generally responded strongly, whereas the response of glial fibrillary acidic protein-positive astrocytes appeared to be more variable. Importantly, some reactive cells also expressed both microglia and astrocytic markers, thus confounding their origin. Comparison with post-mortem human brain tissue obtained at rapid autopsies suggested that the reactive process is not a consequence of epilepsy.

Nestin‐expressing cell types in the temporal lobe and hippocampus: Morphology, differentiation, and proliferative capacity

“…MCM2 + NEC were present in all zones in epilepsy cases and with significantly higher proliferative fractions of 6.6% compared with 3.3% in controls. This compares to our previous observations of proliferative fractions of 50% at acute injury sites to under 10% in chronic scars (Goc et al, ) suggesting a higher baseline activation of NEC in epilepsy as in previous studies (Verwer et al, ). The regions we noted with higher Nestin + /MCM2 + cell densities were the hippocampal sulcus and PVWM.…”

“…Quantitative studies comparing nestin expression in surgically resected TLE to PM control tissue in culture showed significantly higher levels in epilepsy cases (Verwer et al, 2015). We also demonstrated in our series significantly more NEC in epilepsy than controls.…”

“…Proliferation of NEC types is well recognized at injury sites as part of the normal process of brain repair (Burda, Bernstein, & Sofroniew, ). An in vitro slice culture study in TLE patients following surgery showed an upregulation of NEC in relation to vessels (Verwer et al, ). In our previous study of the time course of NEC proliferation in relation to cortical injuries following depth electrode recordings, cells typically exhibited a radial‐glial like bipolar morphology and a close relationship to new‐formed vessels in acute lesions (Goc et al, ).…”

“…There is evidence that acute seizures can increase nestin expression (Jansson, Wennstrom, Johanson, & Tingstrom, ) and following status epilepticus, nestin + radial glial fibers have been reported (Scorza et al, ). Quantitative studies comparing nestin expression in surgically resected TLE to PM control tissue in culture showed significantly higher levels in epilepsy cases (Verwer et al, ). We also demonstrated in our series significantly more NEC in epilepsy than controls.…”

Alterations in the steroid biosynthetic pathways in the human prefrontal cortex in mood disorders: A post‐mortem study

“…We wondered whether this effect could also be detected in DHEA treated human brain tissue. In principle, cultures of post‐mortem tissue or resected tissue might be used for this purpose. We decided to use post‐mortem brain tissue because cells in this tissue remain alive for long periods in culture, whereas neurons quickly degenerate in resected tissue .…”