Stem cells and differentiated cells differ in their sensitivity to urine in vitro

Area of Science:

• Regenerative Medicine
• Urology
• Tissue Engineering

Background:

• Urinary tract regeneration is a complex challenge in reconstructive urology.
• Cells used in tissue engineering scaffolds are immediately exposed to urine post-implantation.
• Cellular response to urine is critical for successful bladder regeneration.

Purpose of the Study:

• To compare the in vitro sensitivity of three distinct cell lines to urine.
• To evaluate the impact of urine exposure on cell viability, proliferation, and apoptosis/necrosis.

Main Methods:

• Isolation of porcine urothelial cells (UCs), smooth muscle cells (SMCs), and adipose-derived stem cells (ADSCs).
• Assessment of cell viability using MTT assay.
• Real-time proliferation analysis via the xCELLigence system.
• Flow cytometry for apoptosis/necrosis determination after urine exposure.

Main Results:

• ADSCs exhibited the highest sensitivity to urine compared to UCs and SMCs.
• UCs demonstrated greater resistance to urine than SMCs.
• Twenty-four-hour urine incubation reduced viability by ~40% (UCs), ~70% (SMCs), and ~90% (ADSCs).

Conclusions:

• UCs and SMCs are more suitable for urinary bladder regeneration than ADSCs due to higher urine resistance.
• Urine-mediated cytotoxicity mechanisms vary among cell types.
• Mesenchymal stem cells like ADSCs have limitations in bladder regeneration applications, particularly concerning urinary bladder cancer.