Glycerol-Induced Acute Kidney Injury Model
Objective: In vivo assessment of microvesicle effects on morphologic and functional recovery from glycerol-induced acute kidney injury in SCID mice
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Materials3
Software2
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Protocol Steps
Induce acute kidney injury
Glycerol-induced acute kidney injury model established in SCID mice
Note: This is the baseline injury model against which treatments are tested
View evidence from paper
“microvesicles accelerated the morphologic and functional recovery of glycerol-induced AKI in SCID mice”
Administer microvesicles
Mesenchymal stem cell-derived microvesicles administered to injured mice to assess protective effects
Note: Microvesicles require CD44- and β1-integrin-dependent incorporation into tubular cells for biologic action
View evidence from paper
“The biologic action of microvesicles required their CD44- and β1-integrin-dependent incorporation into tubular cells”
Assess morphologic recovery
Evaluate morphologic changes in kidney tissue following microvesicle treatment
Note: Microvesicles accelerated morphologic recovery compared to control
View evidence from paper
“microvesicles accelerated the morphologic and functional recovery of glycerol-induced AKI”
Assess functional recovery
Evaluate functional parameters of kidney following microvesicle treatment
Note: Functional recovery was accelerated by microvesicle treatment similar to MSC treatment
View evidence from paper
“The effect of microvesicles on the recovery of AKI was similar to the effect of human MSCs”
Analyze microvesicle RNA content
Extract and analyze RNA from microvesicles using microarray analysis and quantitative real time PCR
Note: Analysis identified specific subset of cellular mRNA including mRNAs associated with mesenchymal phenotype and control of transcription, proliferation, and immunoregulation
View evidence from paper
“Microarray analysis and quantitative real time PCR of microvesicle-RNA extracts indicate that microvesicles shuttle a specific subset of cellular mRNA”
Test RNA-dependent effects with RNase
Treat microvesicles with RNase to determine if biologic effects are RNA-dependent
Note: RNase treatment abolished protective effects both in vitro and in vivo, confirming RNA-dependent mechanism
View evidence from paper
“RNase abolished the aforementioned effects of microvesicles in vitro and in vivo, suggesting RNA-dependent biologic effects”