Purified human skeletal muscle-derived stem cells enhance the repair and regeneration in the damaged urethra.

Background: Postoperative damage of the urethral rhabdosphincter and nerve-vascular networks is a major complication of radical prostatectomy and generally causes incontinence and/or erectile dysfunction. The human skeletal muscle-derived stem cells, which have a synchronized reconstitution capacity of muscle-nerve-blood vessel units, were applied to this damage. Methods: Cells were enzymatically extracted from the human skeletal muscle, sorted using flow cytometry as CD34+/45- (Sk-34) and CD29+/34-/45- (Sk-DN/29+) fractions, and separately cultured/expanded in appropriate conditions within 2 weeks. Urethral damage was induced by manually removing one-third of the wall of the muscle layer in nude rats. A mixture of expanded Sk-34 and Sk-DN/29+ cells was applied on the damaged portion for the cell transplantation (CT) group. The same amount of media was used for the non-cell transplantation (NT) group. Urethral pressure profile was evaluated via electrical stimulation to assess functional recovery. Cell engraftments and differentiations were detected using immunohistochemistry and immunoelectron microscopy. Expression of angiogenic cytokines was also analyzed using RT-PCR and protein array. Results: At 6 weeks after transplantation, the CT-group showed a significantly higher functional recovery than the NT-group (70.2% and 39.1%, respectively; p

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