Cutaneous Wound Model
Objective: To assess mesenchymal stem cell (MSC) engraftment and localization in cutaneous wounds using in vivo bioluminescent imaging to monitor MSC tropism for inflammatory sites
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Equipment1
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Materials1
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Software1
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Protocol Steps
Create cutaneous wounds
Induce inflammatory insults in mice using two wound types: needle-stick wounds and surgical incision wounds
Note: Wounds serve as inflammatory sites for MSC engraftment assessment
View evidence from paper
“Inflammatory insults investigated included cutaneous needle-stick and surgical incision wounds”
Prepare MSC-ffLuc cells
Prepare mesenchymal stem cells modified to express firefly luciferase for injection
Note: Cells must express firefly luciferase for bioluminescent detection
View evidence from paper
“MSC modified to express firefly luciferase (MSC-ffLuc) were injected into healthy mice or mice bearing inflammatory insults”
Inject MSC-ffLuc systemically
Deliver MSC-ffLuc cells to mice via systemic injection (intravenous or intraperitoneal routes)
Note: Both intravenous and intraperitoneal delivery routes were compared in tumor models
View evidence from paper
“MSC-ffLuc systemically delivered to non-tumor bearing animals initially reside in the lungs, then egress to the liver and spleen”
Monitor MSC localization with bioluminescent imaging
Use noninvasive in vivo bioluminescent imaging to track MSC localization over time in wounded and non-wounded animals
Note: Imaging allows monitoring of MSC dispersion in single animals and concurrent observation of multiple variables
View evidence from paper
“MSC localization was followed with bioluminescent imaging”
Assess MSC engraftment in wounded mice
Determine whether MSC engraft and remain detectable specifically at injured wound sites in wounded animals
Note: Results compared between wounded and non-wounded animals
View evidence from paper
“hMSC in wounded mice engraft and remain detectable only in injured sites”