Targeted intraspinal injections to assess therapies in rodent models of neurological disorders

Autor: Eva L. Feldman, Maegan A. Tabbey, Kevin S. Chen, Stacey A. Sakowski, John M. Hayes, Lisa M. McGinley, Elizabeth S. Bruno, Josh S. Chang, Osama N. Kashlan, Faye E. Mendelson, Karl Johe
Rok vydání: 2019
Předmět:
Zdroj: Nature Protocols. 14:331-349
ISSN: 1750-2799
1754-2189
DOI: 10.1038/s41596-018-0095-5
Popis: Despite decades of research, pharmacological therapies for spinal cord motor pathologies are limited. Alternatives using macromolecular, viral, or cell-based therapies show early promise. However, introducing these substances into the spinal cord, past the blood–brain barrier, without causing injury is challenging. We describe a technique for intraspinal injection targeting the lumbar ventral horn in rodents. This technique preserves motor performance and has a proven track record of translation into phase 1 and 2 clinical trials in amyotrophic lateral sclerosis (ALS) patients. The procedure, in brief, involves exposure of the thoracolumbar spine and dissection of paraspinous muscles over the target vertebrae. Following laminectomy, the spine is affixed to a stereotactic frame, permitting precise and reproducible injection throughout the lumbar spine. We have used this protocol to inject various stem cell types, primarily human spinal stem cells (HSSCs); however, the injection is adaptable to any candidate therapeutic cell, virus, or macromolecule product. In addition to a detailed procedure, we provide stereotactic coordinates that assist in targeting of the lumbar spine and instructional videos. The protocol takes ~2 h per animal. This protocol describes how to inject therapeutic cells, viruses, or other macromolecular products into the mouse or rat lumbar ventral cord, modeling the routes used for administration of therapies currently in use in human clinical trials.
Databáze: OpenAIRE
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