The Differentiation Of Muscle Derived Stem Cells In The Rat Penile Corpus Cavernosum Into Smooth Muscle Cells In Vitro

Objective: To isolate and culture muscle derived stem cells (MDSCs) of the rat penilecorpus cavernosum and detect the expression of stem cell markers. To investigate MDSCsstem cell self-renewal and continued proliferation capacity. And to observe theirdifferentiation potential to smooth muscle cells in definite culture．Method: The penile corpus cavernosum were taken under sterile condition from2-month-old Sprague-Dawley rats, MDSCs Were purified through percoll density gradientcentrifugation and preplate technique. PP6cells were obtained and detected expression ofstem cell markers Sca-1and myoblast marker Desmim by immunofluorescence and Flowcytometry. Counting the number of passaged cells, the growth curve of third, sixth, eightgenerations of passage cells were drawn, and the expression of Sca-1, Desmim wasdetected in these passaged cells. The isolated cells were induced to differentiation intosmooth muscle lineage by coculturing with corpus cavernosal smooth muscle cells(CCSMCs). Immunofluorescence were performed to detect the expression of specifica-SMA.Results: MDSCs were purified through percoll density gradient centrifugation andpreplate technique. MDSCs has small body, lower adherence and well refraction,andappear as globular or fusiforn in the first72hours, then the number，cell density, cellvolume of MDSCs are increased and the cells morphology is changed,appearing as spindle,prismatic after5-7d, in12-14d division growth is obvious,and after14-16d cell fusion isabout80%-90%.Positive immunofluorescence outcomes were found with the stem cellmarkers and myoblast markers. The growth curve showed that the cell doubling period isabout48hours,into the platform during the seventh day in vitro,cell proliferation did notchange significantly until the eighth generation.After2days’ co-cultrue induction, theisolated cells expressed the specific a-SMA.Conclusion: The cells from rat penile corpus cavernosum had the cell phenotype of muscle derived stem cells, which can self-renew and proliferate stably. they could beinduced into smooth muscle cells in vitro. Which give the further evidence that there areendogenous muscle-derived stem cells in the penis.