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Objective: The aim of this work is to develop an organotypic culture of cortical human neurons derived from a contusion tissue obtain from patients that suffered TBI.
Methodology: We used contused brain tissue from 4 TBI patients. Sections between 1,500-2,000 mm were kept in a continuous flow of aCSF 2 ml/min in a mixture of 95% O2 and 5% CO2 for 2, 8 and 14 hours. The initial time (0 hours) was the tissue extraction time. From blocks, sections of 50 mm were obtained and processed for immunocytochemistry to NeuN and MAP2.
Results: The results show that organotypic cultures keep neuron integrity and laminar organization in the cerebral cortex slices from 0 to 2 hours. From this time ahead neuronal morphology and laminar organization is altered especially in neurons located on layers III and V.
Conclusions: Organotypic culture could be maintained from 0-2 hours. Neuronal and laminar integrity could be demonstrated. The model lead to study neuronal behaviour after TBI through different survival times. Laminar selective vulnerability was demonstrated for layers III and V.
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