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Nat Protoc. 2019 Feb;14(2):331-349. doi: 10.1038/s41596-018-0095-5.

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

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Department of Neurosurgery, University of Michigan, Ann Arbor, MI, USA.
Department of Neurology, University of Michigan, Ann Arbor, MI, USA.
Neuralstem, Germantown, MD, USA.
Department of Neurology, University of Michigan, Ann Arbor, MI, USA.


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.


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