An organotypic brain slice preparation from adult patients with temporal lobe epilepsy

J Neurosci Methods. 2014 Sep 30;235:234-44. doi: 10.1016/j.jneumeth.2014.07.009. Epub 2014 Jul 23.

Abstract

Background: A long-term in vitro preparation of diseased brain tissue would facilitate work on human pathologies. Organotypic tissue cultures retain an appropriate neuronal form, spatial arrangement, connectivity and electrical activity over several weeks. However, they are typically prepared with tissue from immature animals. In work using tissue from adult animals or humans, survival times longer than a few days have not been reported and it is not clear that pathological neuronal activities are retained.

New method: We modified tissue preparation procedures and used a defined culture medium to make organotypic cultures of temporal lobe tissue obtained after operations on adult patients with pharmaco-resistant mesial temporal lobe epilepsies.

Results: Organototypic culture preparation and maintenance techniques were judged on criteria of morphology and the generation of epileptiform activities. Short-duration (30-100 ms) interictal-like population activities were initiated spontaneously in either the subiculum, dentate gyrus or the CA2/CA3 region, but not the cortex, for up to 3-4 weeks in culture. Ictal-like discharges, of duration greater than 10s, were induced by convulsants. Epileptiform activities were modulated by both glutamatergic and GABAergic receptor antagonists.

Comparison with existing methods: Our methods now permit the maintenance in organotypic culture of epileptic adult human tissue, generating appropriate epileptiform activity over 3-4 weeks.

Conclusions: We have shown that characteristic morphology and pathological activities are maintained in organotypic cultures of adult human tissue. These cultures should permit studies on the effects of prolonged drug treatments and long-term procedures such as viral transduction.

Keywords: Defined medium; Epilepsy; Hippocampus; Human; Organotypic culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Culture Media
  • Epilepsy, Temporal Lobe / drug therapy
  • Epilepsy, Temporal Lobe / pathology
  • Epilepsy, Temporal Lobe / physiopathology*
  • Epilepsy, Temporal Lobe / surgery*
  • Excitatory Amino Acid Antagonists / pharmacology
  • Female
  • GABA Antagonists / pharmacology
  • GABAergic Neurons / drug effects
  • GABAergic Neurons / pathology
  • GABAergic Neurons / physiology
  • Humans
  • Immunohistochemistry
  • Male
  • Microelectrodes
  • Microscopy, Electron
  • Middle Aged
  • Patch-Clamp Techniques
  • Receptors, Glutamate / metabolism
  • Temporal Lobe / drug effects
  • Temporal Lobe / pathology
  • Temporal Lobe / physiopathology*
  • Temporal Lobe / surgery*
  • Time Factors
  • Tissue Culture Techniques / methods*
  • Young Adult

Substances

  • Culture Media
  • Excitatory Amino Acid Antagonists
  • GABA Antagonists
  • Receptors, Glutamate