Organotypic human neuronal culture derived from traumatic brain injury.

Abstract

Introduction: Traumatic brain injury is a global medical problem whose survivors may show disability and neurological or psychiatric sequelae. In the last few years the knowledge of physiopathological mechanisms of TBI has increase but still it is not entirely known. For this reason the research has turn over in one´s mind in new strategies to study this pathology looking for neuroprotection.
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.

Authors

• David Riascos Universidad del Valle
• Francisco Guzmán Universidad del Valle
• Efraín Buriticá Universidad del Valle
• Mauricio Palacios Universidad del Valle
• Martha Isabel Escobar Universidad del Valle
• Hernán Pimienta Universidad del Valle