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Transdifferentiation of rat fetal brain stem cells into penile smooth muscle cells
Yunseob Song1, Neal Mehta, Bryant Sheh
1Department of Urology, Soonchunhyang University College of Medicine, Seoul, Korea.
Objectives:
To evaluate whether rat fetal brain stem cells can be induced to acquire cell fates outside the nervous system, hypothesising that cell-based replacement therapy with stem cells can aid in the regeneration of penile smooth musculature and might help to attenuate organic erectile dysfunction (ED), as the degeneration of penile smooth muscle cells leading to subsequent impairment of function is important in organic ED.
Materials And Methods:
Fetal brain stem cells (FBSCs) from embryonal 12-day Fisher 344 rats were isolated and characterized. For in vitro studies, undifferentiated FBSCs were cultured for 21 days in either N2 media (control) or N2 media conditioned in rat penile smooth muscle cell culture. These were then subjected to immunocytochemistry for specific markers of neural stem cells (nestin) and penile smooth muscle cells, i.e. alpha-smooth muscle actin (alphaSMA), penis-specific myosin light chain (MLC) desmin, calponin, vimentin, phosphodiesterase-5 (PDE5) and connexin. For in vivo studies, male adult Fisher 344 rats had an intracavernous injection with saline (five rats, control) or FBSCs that were labelled genetically by an expression construct for green fluorescent protein (GFP, nine rats, experimental) and maintained for 6 weeks. The rats were then killed and penile tissue was harvested and subjected to immunocytochemistry for markers of neural stem cells, smooth muscle cells, and sinusoidal endothelium (vascular endothelial growth factor, VEGF).
Results:
Undifferentiated cells exposed to N2 media continued to maintain the characteristic morphological and protein marker features of FBSCs, while the cells exposed to the conditioned media acquired the morphological features of smooth muscle cells. In addition, the differentiated cells (30-40%) expressed smooth muscle markers. Rats implanted with FBSCs had cells that showed double-labelling for GFP/alphaSMA, GFP/calponin and GFP/VEGF. The control group had no evidence of such double-labelling.
Conclusions:
These results suggest the transdifferentiation of FBSCs into penile smooth muscle cells. Such transdifferentiated cells showed long-term survival when injected into the cavernous tissue, thus raising the possibility of a novel therapeutic option for organic ED.
