Home > Browse Issues > Vol.26 No.6
Establishment of Organotypic Culture Model of Spinal Cord for Amyotrophic Lateral Sclerosis
Wei-Gang Liu, Xiao-Juan Wang1, Xiang-Jian Xiao2, Zheng Ma, Xue-Qin Song, Li-Qin Wang, Chun-Yan Li*
Department of Neurology, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, China; 1Department of Neurology, Beijing Tongren Hospital, Beijing 100730, China; 2Department of Neurology, the People's Hospital of Hebei Province, Shijiazhuan
Abstract: In this study， we developed a cultured organotypic spinal cord model of selective motor neuron death for amyotrophic lateral sclerosis (ALS)， using threohydroxyaspartate (THA)， a inhibitor of glutamate transport. Cultured organotypic spinal cord slices were prepared from lumbar spinal cords of 8-day-old rat pups. Various concentrations of THA were continuously added into the culture medium. Ventral a-motor neuron survival was evaluated by culture morphology and SMI-32 immunohistochemistry staining. Interneurons in dorsal horn were identified by calretinin immunohistochemistry staining. Glutamate (Glu) and lactate dehydrogenase (LDH) levels in culture medium were measured. The results showed that the spinal cord slices in the control group could maintain excellent organotypic cellular organization and a stable population of ventral SMI-32 positive a-motor neurons. THA could produce a slow dose-dependent loss of SMI-32 positive a-motor neurons and a elevation of Glu ， LDH levels in culture medium. After the slices cultured 4 weeks in vitro， 100 mmol/L THA resulted in the loss of SMI-32 positive a-motor neurons， increase of Glu level and no significant change of inter-neurons in the dorsal horn， which could be acted as an effective organotypic culture model of ALS.
CN
EN