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

Abstract

Urinary tract regeneration using tissue engineering is one of the most challenging issues in the field of reconstructive urology. Cells seeded on scaffold are exposed to urine immediately after the implantation. The outcome of urinary bladder regeneration is depended on the ability of these cells to survive, proliferate, and regenerate. The aim of this study was to compare a sensitivity of three different cell lines to urine in vitro. Three different cell lines were isolated from porcine bladder (urothelial cells, UCs and smooth muscle cells, SMCs) and adipose tissue (adipose-derived stem cells, ADSCs). Cell viability (MTT assay), proliferation (real-time cell analysis using xCELLigence system) and apoptosis/necrosis (flow cytometry) were analyzed after exposition to urine. ADSCs were the most sensitive to urine compared to two other tested cell lines. Among the bladder cell lines the UCs were more resistant to urine than SMCs. Twenty four hour incubation of UCs, SMCs, and ADSCs with urine lead to ∼40%, ∼70%, and ∼90% reduction of their viability, respectively. The mechanism of urine mediated cytotoxicity differed depending on the tested cell type. Urothelial and SMCs seems to be more suitable for urinary bladder regeneration compared to mesenchymal stem cells, however, these cells have limited application especially in the case of urinary bladder cancer.

Understanding the role of mesenchymal stem cells in urinary bladder regeneration—a preclinical study on a porcine model

“…Microenvironment of the regenerating bladder wall is highly cytotoxic due to presence of urine which was demonstrated to kill predominantly undifferentiated stem cells. Even short 24 h ADSC incubation with urine reduced the colony viability by 90% [ 16 ]. Taking low chances for long ADSC survival might only provide temporary trophic boost for regenerating bladder tissue.…”

Tissue Engineering and Its Potential to Reduce Prostate Cancer Treatment Sequelae—Narrative Review

“…Furthermore, the persistence of urine within the healing anastomosis is an often underestimated factor that may have a negative effect on final anastomosis remodeling. It was demonstrated that urine has a cytotoxic effect on muscles precursor cells participating in urinary tract wall regeneration (30). The primary approach of tissue engineering should aim to transform heeling pattern within the vesicourethral anastomosis and adjacent tissues.…”

Urine as a Main Effector in Urological Tissue Engineering—A Double-Edged Sword

“…Available evidence indicates that urothelial cells are more resistant to urine as compared to underling smooth muscle cells. In all cell lines, the toxic effect of urine has been confirmed by a real-time analysis, and simple urine addition has been shown to result in a fast decline in cell proliferation [17]. In comparison to Urothelial Cells (UCs) that were substantially resistant, adipose-derived stromal cells (ADSCs) are the most sensitive cell type [17].…”

“…Bioengineered urothelium would present a valuable tool for urethral replacement, as it is resistant to the toxic effects of urine [17,20]. Presently, limited in vitro models of urothelium exist for different urologic diseases within the lower urinary tract to be treated by medications.…”