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A Method to Isolate Pericytes From the Mouse Urinary Bladder for the Study of Diabetic Bladder Dysfunction
Min-Ji Choi1, Nguyen Nhat Minh1, Jiyeon Ock1
1National Research Center for Sexual Medicine and Department of Urology, Inha University College of Medicine, Incheon, Korea.
International Neurourology Journal
|January 6, 2021
Summary
Researchers isolated mouse urinary bladder pericytes and developed a model for diabetic bladder dysfunction. High glucose impaired pericyte function, reducing tube formation and proliferation while increasing apoptosis and oxidative stress.
Area of Science:
- Vascular Biology
- Cell Biology
- Diabetology
Background:
- Pericytes are crucial for microvessel integrity and function.
- Pericyte dysfunction is implicated in diabetic complications like retinopathy and erectile dysfunction.
- Diabetic bladder dysfunction is a significant health concern with limited therapeutic options.
Purpose of the Study:
- To establish a method for isolating and culturing pericytes from the mouse urinary bladder.
- To develop an in vitro model of diabetic bladder dysfunction using these pericytes.
- To investigate the effects of high glucose on isolated bladder pericyte function.
Main Methods:
- Meticulous removal of the urothelial layer to isolate submucosa and detrusor muscle.
- Culture of fragmented tissues on collagen I-coated plates under normal or high glucose.
- Assessment of tube formation, cell proliferation, apoptosis (TUNEL assay), and superoxide anion production (hydroethidine staining).
Main Results:
- Successful isolation of high-purity mouse urinary bladder pericytes, confirmed by specific marker expression (PDGFR-β, NG2).
- High glucose significantly reduced pericyte tube formation and proliferation.
- High glucose exposure led to increased superoxide production and apoptosis in bladder pericytes.
Conclusions:
- This study presents the first isolation and culture of pericytes from the mouse urinary bladder.
- The developed in vitro model effectively mimics diabetic bladder dysfunction at the cellular level.
- This model serves as a valuable platform for screening therapeutic agents targeting pericyte function in diabetic bladder complications.
