Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury methods
Aim. Evidence-backed execution summary for Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury methods from Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury.
Show snapshot details
On this page
This experiment, in seven questions
Jump straight to the part of the recipe you need. Data and provenance labels stay close to the action they support.
Shopping and prep list
What do I need before I start?
rat
Subject model for the experiment.
- Use
- confirm full cohort details in the source paper
Methods
reagent used in the protocol.
- Use
- MSC expansion was performed according to previously described methods. Briefly, bone marrow from adult male rats was mechanically harvested by flushing the cavity of the femurs with PBS, and the cells were washed and suspended in culture medium. Three days later, cells that tightly adhered to the plastic flasks were...
Results
reagent used in the protocol.
- Use
- Exosomes accumulate as intraluminal vesicles inside multivesicular bodies,,,, while microvesicles (size 100∼1000 nm) are small, plasma-membrane-derived particles that are released into the extracellular environment by the outward budding and fission of the plasma membrane.,, A precise and clear distinctio...
Methods
Wistar rats were subjected to TBI followed by tail vein injection of 100 µg protein of exosomes derived from MSCs or an equal volume of vehicle phosphate-buffered saline (n = 8/group) 24 hours later. To evaluate cognitive and sensorimotor functional recovery, the modified Morris water maze, neurological severit...
- Use
- Wistar rats were subjected to TBI followed by tail vein injection of 100 µg protein of exosomes derived from MSCs or an equal volume of vehicle phosphate-buffered saline (n = 8/group) 24 hours later. To evaluate cognitive and sensorimotor functional recovery, the modified Morris water maze, neurological severit...
Methods
All experimental procedures were approved by the Henry Ford Health System Institutional Animal Care and Use Committee. To prevent potential biases of performance and detection, the persons who performed the experiments, collected data, and assessed outcome were blinded throughout the course of the experiments and we...
- Use
- All experimental procedures were approved by the Henry Ford Health System Institutional Animal Care and Use Committee. To prevent potential biases of performance and detection, the persons who performed the experiments, collected data, and assessed outcome were blinded throughout the course of the experiments and we...
Methods
MSC expansion was performed according to previously described methods. Briefly, bone marrow from adult male rats was mechanically harvested by flushing the cavity of the femurs with PBS, and the cells were washed and suspended in culture medium. Three days later, cells that tightly adhered to the plastic flasks were...
- Use
- MSC expansion was performed according to previously described methods. Briefly, bone marrow from adult male rats was mechanically harvested by flushing the cavity of the femurs with PBS, and the cells were washed and suspended in culture medium. Three days later, cells that tightly adhered to the plastic flasks were...
Morris Water Maze (MWM) Test
To measure spatial learning impairments, an updated version of the MWM test was used. The procedure was modified from previous versions, and has been used for spatial memory assessment in rodents with brain injury. The MWM test was performed monthly postinjury. At each testing interval, animals were tested with 4 tr...
- Use
- To measure spatial learning impairments, an updated version of the MWM test was used. The procedure was modified from previous versions, and has been used for spatial memory assessment in rodents with brain injury. The MWM test was performed monthly postinjury. At each testing interval, animals were tested with 4 tr...
Results
Exosomes accumulate as intraluminal vesicles inside multivesicular bodies,,,, while microvesicles (size 100∼1000 nm) are small, plasma-membrane-derived particles that are released into the extracellular environment by the outward budding and fission of the plasma membrane.,, A precise and clear distinctio...
- Use
- Exosomes accumulate as intraluminal vesicles inside multivesicular bodies,,,, while microvesicles (size 100∼1000 nm) are small, plasma-membrane-derived particles that are released into the extracellular environment by the outward budding and fission of the plasma membrane.,, A precise and clear distinctio...
Before you run
What should be confirmed before execution?
First confirmation
Equipment is listed but no product mappings are linked.
Confirm before execution
This page is backed by a publishable Replication Data Ledger package with zero critical source-verification issues.
Confirm before execution
Open the source paper before finalizing run-specific details.
Procurement checkpoint
Use source-stated vendors where present. Treat mapped products as sourcing options unless the page marks an exact source match.
Open quote workflowStep-by-step procedure
What do I do, in order?
Methods
Wistar rats were subjected to TBI followed by tail vein injection of 100 µg protein of exosomes derived from MSCs or an equal volume of vehicle phosphate-buffered saline (n = 8/group) 24 hours later. To evaluate cognitive and sensorimotor functional recovery, the modified Morris water maze, neurological severity score and footfault tests were performed. Animals were sacrificed at 35 days after TBI. Histopathological and immunohistochemical analyses were performed for measurements of lesion volume, neurovascular remodeling (angiogenesis and neurogenesis), and neuroinflammation.
Methods
MSC expansion was performed according to previously described methods. Briefly, bone marrow from adult male rats was mechanically harvested by flushing the cavity of the femurs with PBS, and the cells were washed and suspended in culture medium. Three days later, cells that tightly adhered to the plastic flasks were considered as P0 MSCs. MSCs were conventionally cultured with a modified MEM medium (Hyclone, Logan, UT, USA) containing 20% fetal bovine serum (Gibson Laboratory, Grand Island, NY, USA) and penicillin-streptomycin on 75cm tissue culture flasks (Corning, St Louis, MO, USA). For the exosome isolation, conventional culture medium was replaced with an exosome-depleted fetal bovine serum-contained (EXO-FBS-250 A-1, System Biosciences, Mountain View, CA, USA) medium when the cells reached 60% to 80% confluence, and the MSCs were cultured for an additional 48 hours. The me...
Morris Water Maze (MWM) Test
To measure spatial learning impairments, an updated version of the MWM test was used. The procedure was modified from previous versions, and has been used for spatial memory assessment in rodents with brain injury. The MWM test was performed monthly postinjury. At each testing interval, animals were tested with 4 trials per day for 5 consecutive days on Day 31-35 after mTBI. A blue swimming pool (1.8 m in diameter) was located in a large room, where there were many clues external to the maze (e.g., pictures on the walls, lamps and a camera on the ceiling); these were visible from the pool and presumably used by the rats for spatial orientation. The position of the cues remained unchanged throughout the experiment. Data collection was automated using the HVS Image 2020 Plus Tracking System (US HVS Image, San Diego, CA), as described previously. For data collection, the swimming pool wa...
Measurement outputs
What raw and processed outputs should exist?
Wistar rats were subjected to TBI followed by tail vein injection of 100 µg protein of exosomes derived from MSCs or an equal volume of vehicle phosphate-buffered saline (n...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
All experimental procedures were approved by the Henry Ford Health System Institutional Animal Care and Use Committee. To prevent potential biases of performance and detection,...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
To measure spatial learning impairments, an updated version of the MWM test was used. The procedure was modified from previous versions, and has been used for spatial memory ass...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Exosomes accumulate as intraluminal vesicles inside multivesicular bodies,,,, while microvesicles (size 100∼1000 nm) are small, plasma-membrane-derived particles that a...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Analysis plan
How should the outputs become interpretable results?
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
inferred from protocolPreprocessing / cleaning
To measure spatial learning impairments, an updated version of the MWM test was used.
from paperScoring or quantification
Quantify the primary readouts for this experiment: Wistar rats were subjected to TBI followed by tail vein injection of 100 µg protein of exosomes derived from MSCs or an equal volume of vehicle phosphate-buffered saline (n...; All experimental procedures were approved by the Henry Ford Health System Institutional Animal Care and Use Committee. To prevent potential biases of performance and detection,...; To measure spatial learning impairments, an updated version of the MWM test was used. The procedure was modified from previous versions, and has been used for spatial memory ass...; Exosomes accumulate as intraluminal vesicles inside multivesicular bodies,,,, while microvesicles (size 100∼1000 nm) are small, plasma-membrane-derived particles that a....
from paperStatistical comparison
To measure spatial learning impairments, an updated version of the MWM test was used. The procedure was modified from previous versions, and has been used for spatial memory ass...
from paperReporting output
Report representative outputs alongside summary comparisons for Wistar rats were subjected to TBI followed by tail vein injection of 100 µg protein of exosomes derived from MSCs or an equal volume of vehicle phosphate-buffered saline (n..., All experimental procedures were approved by the Henry Ford Health System Institutional Animal Care and Use Committee. To prevent potential biases of performance and detection,..., To measure spatial learning impairments, an updated version of the MWM test was used. The procedure was modified from previous versions, and has been used for spatial memory ass..., Exosomes accumulate as intraluminal vesicles inside multivesicular bodies,,,, while microvesicles (size 100∼1000 nm) are small, plasma-membrane-derived particles that a....
inferred from protocolStructured statistical methods
To measure spatial learning impairments, an updated version of the MWM test was used. The procedure was modified from previous versions, and has been used for spatial memory ass...
source structuredSource and audit
What supports the facts on this page?
Evidence quotes (3)
Wistar rats were subjected to TBI followed by tail vein injection of 100 µg protein of exosomes derived from MSCs or an equal volume of vehicle phosphate-buffered saline (n = 8/group) 24 hours later. To evaluate cognitive and sensorimotor functional recovery, the modified Morris water maze, neurological severity score and footfault tests were performed. Animals were sacrificed at 35 days after TBI. Histopathological and immunohistochemical analyses were performed for measurements of lesion volume, neurovascular remodeling (angiogenesis and neurogenesis), and neuroinflammation.
MSC expansion was performed according to previously described methods. Briefly, bone marrow from adult male rats was mechanically harvested by flushing the cavity of the femurs with PBS, and the cells were washed and suspended in culture medium. Three days later, cells that tightly adhered to the plastic flasks were considered as P0 MSCs. MSCs were conventionally cultured with a modified MEM medium (Hyclone, Logan, UT, USA) containing 20% fetal bovine serum (Gibson Laboratory, Grand Island, NY, USA) and penicillin-streptomycin on 75cm tissue culture flasks (Corning, St Louis, MO, USA). For the exosome isolation, conventional culture medium was replaced with an exosome-depleted fetal bovine serum-contained (EXO-FBS-250 A-1, System Biosciences, Mountain View, CA, USA) medium when the cells reached 60% to 80% confluence, and the MSCs were cultured for an additional 48 hours. The media were then collected and exosomes were isolated by ExoQuick exosome isolation method according to the manufacture's instruction. Briefly, ExoQuick-TC (2.5 ml) was added to 10 ml of media, incubated 12 hours at 4°C, and centrifuged at 1500 × g for 30 min to obtain pelleted exosomes. The...
To measure spatial learning impairments, an updated version of the MWM test was used. The procedure was modified from previous versions, and has been used for spatial memory assessment in rodents with brain injury. The MWM test was performed monthly postinjury. At each testing interval, animals were tested with 4 trials per day for 5 consecutive days on Day 31-35 after mTBI. A blue swimming pool (1.8 m in diameter) was located in a large room, where there were many clues external to the maze (e.g., pictures on the walls, lamps and a camera on the ceiling); these were visible from the pool and presumably used by the rats for spatial orientation. The position of the cues remained unchanged throughout the experiment. Data collection was automated using the HVS Image 2020 Plus Tracking System (US HVS Image, San Diego, CA), as described previously. For data collection, the swimming pool was subdivided into four equal quadrants formed by imaging lines. At the start of each trial, the rat was placed at one of four fixed starting points, randomly facing toward a wall (designated North, South, East and West) and allowed to swim for 90 seconds or until it found the platform which was tran...
Machine-readable layer
[
{
"@context": "https://schema.org",
"@type": "HowTo",
"name": "Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury methods",
"description": "Evidence-backed execution summary for Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury methods from Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury.",
"totalTime": "PT4320M",
"step": [
{
"@type": "HowToStep",
"position": 1,
"name": "Methods",
"text": "Wistar rats were subjected to TBI followed by tail vein injection of 100 µg protein of exosomes derived from MSCs or an equal volume of vehicle phosphate-buffered saline (n = 8/group) 24 hours later. To evaluate cognitive and sensorimotor functional recovery, the modified Morris water maze, neurological severity score and footfault tests were performed. Animals were sacrificed at 35 days after TBI. Histopathological and immunohistochemical analyses were performed for measurements of lesion volume, neurovascular remodeling (angiogenesis and neurogenesis), and neuroinflammation."
},
{
"@type": "HowToStep",
"position": 2,
"name": "Methods",
"text": "MSC expansion was performed according to previously described methods. Briefly, bone marrow from adult male rats was mechanically harvested by flushing the cavity of the femurs with PBS, and the cells were washed and suspended in culture medium. Three days later, cells that tightly adhered to the plastic flasks were considered as P0 MSCs. MSCs were conventionally cultured with a modified MEM medium (Hyclone, Logan, UT, USA) containing 20% fetal bovine serum (Gibson Laboratory, Grand Island, NY, USA) and penicillin-streptomycin on 75cm tissue culture flasks (Corning, St Louis, MO, USA). For the exosome isolation, conventional culture medium was replaced with an exosome-depleted fetal bovine serum-contained (EXO-FBS-250 A-1, System Biosciences, Mountain View, CA, USA) medium when the cells reached 60% to 80% confluence, and the MSCs were cultured for an additional 48 hours. The me..."
},
{
"@type": "HowToStep",
"position": 3,
"name": "Morris Water Maze (MWM) Test",
"text": "To measure spatial learning impairments, an updated version of the MWM test was used. The procedure was modified from previous versions, and has been used for spatial memory assessment in rodents with brain injury. The MWM test was performed monthly postinjury. At each testing interval, animals were tested with 4 trials per day for 5 consecutive days on Day 31-35 after mTBI. A blue swimming pool (1.8 m in diameter) was located in a large room, where there were many clues external to the maze (e.g., pictures on the walls, lamps and a camera on the ceiling); these were visible from the pool and presumably used by the rats for spatial orientation. The position of the cues remained unchanged throughout the experiment. Data collection was automated using the HVS Image 2020 Plus Tracking System (US HVS Image, San Diego, CA), as described previously. For data collection, the swimming pool wa..."
}
],
"tool": [
{
"@type": "HowToTool",
"name": "Methods"
},
{
"@type": "HowToTool",
"name": "Methods"
},
{
"@type": "HowToTool",
"name": "Methods"
},
{
"@type": "HowToTool",
"name": "Morris Water Maze (MWM) Test"
},
{
"@type": "HowToTool",
"name": "Results"
}
],
"supply": [
{
"@type": "HowToSupply",
"name": "Methods"
},
{
"@type": "HowToSupply",
"name": "Results"
}
],
"isBasedOn": {
"@type": "ScholarlyArticle",
"headline": "Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury",
"datePublished": "2015",
"author": [
{
"@type": "Person",
"name": "Yanlu Zhang"
},
{
"@type": "Person",
"name": "Michael Chopp"
},
{
"@type": "Person",
"name": "Yuling Meng"
},
{
"@type": "Person",
"name": "Mark Katakowski"
},
{
"@type": "Person",
"name": "Hongqi Xin"
},
{
"@type": "Person",
"name": "Asim Mahmood"
},
{
"@type": "Person",
"name": "Ye Xiong"
}
],
"identifier": "10.3171/2014.11.jns14770"
}
},
{
"@context": "https://schema.org",
"@type": "BreadcrumbList",
"itemListElement": [
{
"@type": "ListItem",
"position": 1,
"name": "Experiments",
"item": "https://replicatescience.com/experiments"
},
{
"@type": "ListItem",
"position": 2,
"name": "Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury methods",
"item": "https://replicatescience.com/experiments/effect-of-exosomes-derived-from-multipluripotent-mesenchymal-stromal-cells-on-functional-recovery-and-neurovascular-plasticity-in-rats-after-traumatic-brain-injury-methods-yanlu-zh/effect-of-exosomes-derived-from-multipluripotent-mesenchymal-stromal-cells-on-functional-recovery-an-mlpgwc9i"
}
]
}
]