Source Paper
Source Paper
Hidenori Suzuki, Christopher S. Ahuja, Ryan P. Salewski, Lijun Li, Kajana Satkunendrarajah et al.
PLoS ONE • 2017
Traumatic spinal cord injuries (SCIs) affect millions of people worldwide; the majority of whom are in the chronic phase of their injury. Unfortunately, most current treatments target the acute/subacute injury phase as the microenvironment of chronically injured cord consists of a well-established glial scar with inhibitory chondroitin sulfate proteoglycans (CSPGs) which acts as a potent barrier to regeneration. It has been shown that CSPGs can be degraded in vivo by intrathecal Chondroitinase ABC (ChABC) to produce a more permissive environment for regeneration by endogenous cells or transplanted neural stem cells (NSCs) in the subacute phase of injury. Using a translationally-relevant clip-contusion model of cervical spinal cord injury in mice we sought to determine if ChABC pretreatment could modify the harsh chronic microenvironment to enhance subsequent regeneration by induced pluripotent stem cell-derived NSCs (iPS-NSC). Seven weeks after injury-during the chronic phase-we delivered ChABC by intrathecal osmotic pump for one week followed by intraparenchymal iPS-NSC transplant rostral and caudal to the injury epicenter. ChABC administration reduced chronic-injury scar and resulted in significantly improved iPSC-NSC survival with clear differentiation into all three neuroglial lineages. Neurons derived from transplanted cells also formed functional synapses with host circuits on patch clamp analysis. Furthermore, the combined treatment led to recovery in key functional muscle groups including forelimb grip strength and measures of forelimb/hindlimb locomotion assessed by Catwalk. This represents important proof-of-concept data that the chronically injured spinal cord can be 'unlocked' by ChABC pretreatment to produce a microenvironment conducive to regenerative iPS-NSC therapy.
Objective: Assessment of forelimb grip strength to measure motor function recovery in mice following spinal cord injury
This is a Grip Strength Measurement protocol using mouse as the model organism. The procedure involves 11 procedural steps, 2 equipment items, 6 materials. Extracted from a 2017 paper published in PLoS ONE.
Model and subjects
mouse • C57BL/6 • female • 8-10 weeks old • 15 to 20g • 70
Study window
~16 week study window
Core workflow
Animal acquisition and housing • Baseline grip strength measurement • Randomization into experimental groups
Primary readouts
Key equipment and reagents
Verified items
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Obtain 70 adult female wild-type C57BL/6 mice aged 8-10 weeks weighing 15-20g from Jackson Laboratories
“A total of 70 adult (8–10 weeks old), female, wild-type C57BL/6 mice (15 to 20g) from Jackson Laboratories”
Measure grip strength at baseline (6 weeks post-injury) before randomization into treatment groups
Note: No statistically significant differences between groups at this timepoint
“Grip strength measurements at six weeks were not statistically different between groups (ANOVA; p = 0.871)”
Randomize 60 injured mice into four experimental groups: control, ChABC, iPS-NSCs, and ChABC + iPS-NSCs
“Six weeks after injury, mice were randomized into four experimental groups (control, ChABC, iPS-NSCs, & ChABC + iPS-NSCs)”
Implant mini-osmotic pump rostral to the injury site under anesthesia
“Seven weeks post-injury, a mini-osmotic pump was implanted rostral to the injury site”
Deliver either ChABC in aCSF or aCSF alone via implanted mini-osmotic pump
“The pump delivered either Chondroitinase ABC (ChABC) in artificial cerebral spinal fluid (aCSF) or aCSF alone”
Remove mini-osmotic pump after one week of treatment delivery
“One week later the pump was explanted”
Transplant iPS-NSCs into spinal cord parenchyma at four sites (two rostral, two caudal), delivering 50,000 cells in 1 µL per site
“induced pluripotent stem cell-derived neural stem cells (iPS-NSC) were transplanted into the cord parenchyma at two rostral and two caudal sites. 50,000 cells in 1 µL were delivered to each site”
Administer Cyclosporine A immunosuppression starting 2 days prior to cell transplantation
“Cyclosporine A immunosuppression was delivered from 2 days prior to transplantation until the end of the study”
Perform behavioral assessments including grip strength measurements throughout the study period
“Behavioral assessments were performed across all 16 weeks of the study”
Conduct electrophysiologic assessments at end of study
“At 16 weeks electrophysiologic assessments were completed”
Perform whole-animal perfusion, fixation, and immunohistochemical analyses
“followed by whole-animal perfusion, fixation and immunohistochemical analyses”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
Assessment of forelimb grip strength to measure motor function recovery in mice following spinal cord injury
Objective
Assessment of forelimb grip strength to measure motor function recovery in mice following spinal cord injury
Subjects
From papermouse • C57BL/6 • female • 8-10 weeks old • 15 to 20g
Sample count
From paper70
Cohort notes
From paperAdult wild-type mice
Animal acquisition and housing
Baseline grip strength measurement
Randomization into experimental groups
Mini-osmotic pump implantation
Forelimb grip strength
From paperANOVA statistical analysis performed on grip strength measurements
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Motor function recovery
From paperANOVA statistical analysis performed on grip strength measurements
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Electrophysiologic measures
From paperANOVA statistical analysis performed on grip strength measurements
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Immunohistochemical findings
From paperANOVA statistical analysis performed on grip strength measurements
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Forelimb grip strength
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Motor function recovery
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Electrophysiologic measures
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Immunohistochemical findings
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
Preprocessing / cleaning
ANOVA statistical analysis performed on grip strength measurements
Scoring or quantification
Quantify the primary readouts for this experiment: Forelimb grip strength; Motor function recovery; Electrophysiologic measures; Immunohistochemical findings.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Forelimb grip strength, Motor function recovery, Electrophysiologic measures, Immunohistochemical findings.
Source links and direct wording from the methods section for validation and deeper review.
Citation
Hidenori Suzuki et al. (2017). Neural stem cell mediated recovery is enhanced by Chondroitinase ABC pretreatment in chronic cervical spinal cord injury. PLoS ONE
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Steps
11
Evidence Quotes
19
Protocol Items
8
Linked Products
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Canonical Sync
Pending
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Steps
11
Evidence
19
Specific Products
1/1
Canonical Sync
Pending
What this score means
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