title: Epileptogenesis in organotypic hippocampal cultures has limited dependence on culture medium composition.
abstract: Rodent organotypic hippocampal cultures spontaneously develop epileptiform activity after approximately 2 weeks in vitro and are increasingly used as a model of chronic post-traumatic epilepsy. However, organotypic cultures are maintained in an artificial environment (culture medium), which contains electrolytes, glucose, amino acids and other components that are not present at the same concentrations in cerebrospinal fluid (CSF). Therefore, it is possible that epileptogenesis in organotypic cultures is driven by these components. We examined the influence of medium composition on epileptogenesis. Epileptogenesis was evaluated by measurements of lactate and lactate dehydrogenase (LDH) levels (biomarkers of ictal activity and cell death, respectively) in spent culture media, immunohistochemistry and automated 3-D cell counts, and extracellular recordings from CA3 regions. Changes in culture medium components moderately influenced lactate and LDH levels as well as electrographic seizure burden and cell death. However, epileptogenesis occurred in any culture medium that was capable of supporting neural survival. We conclude that medium composition is unlikely to be the cause of epileptogenesis in the organotypic hippocampal culture model of chronic post-traumatic epilepsy.
authors.affiliation: Department of Electrical and Computer Engineering, Lehigh University, Bethlehem, Pennsylvania, United States of America.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Neurology, Massachusetts General Hospital, Boston, Massachusetts, United States of America.|Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: Yero
authors.initials: Y
authors.lastname: Saponjian
authors.affiliation: Bioengineering Program, Lehigh University, Bethlehem, Pennsylvania, United States of America.
authors.forename: Mark M
authors.initials: MM
authors.lastname: Mahoney
authors.affiliation: Department of Neurology, Massachusetts General Hospital, Boston, Massachusetts, United States of America.|Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Staley
authors.affiliation: Department of Electrical and Computer Engineering, Lehigh University, Bethlehem, Pennsylvania, United States of America.|Bioengineering Program, Lehigh University, Bethlehem, Pennsylvania, United States of America.
authors.forename: Yevgeny
authors.identifier: http://orcid.org/0000-0001-7539-601X
authors.initials: Y
authors.lastname: Berdichevsky
mesh_terms: D000818:Animals; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D004827:Epilepsy / Q000503:physiopathology*; D006624:Hippocampus / Q000187:drug effects* / Q000503:physiopathology; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D019344:Lactic Acid / Q000032:analysis; D009474:Neurons / Q000187:drug effects* / Q000502:physiology; D009924:Organ Culture Techniques / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D003470:Culture Media; D019344:Lactic Acid; D007770:L-Lactate Dehydrogenase