Abstract:

BACKGROUND: Previous studies have confirmed that stem cells with stem cell markers and muscle-derived stem cell markers were detected in rat penile corpus cavernosum.
OBJECTIVE: To perform isolation and phenotype identification of muscle-derived stem cells in rat penile corpus cavernosum on the basis of previous study.
METHODS: The penile corpus cavernosum from 2-month-old Sprague-Dawley rats was digested and dissociated by type Ⅰ collagenase and trypsin. Cells were primarily purified by the Preplate differential adhesion technique. PP1-PP6 adhered cells were obtained and the expressions of stem cell markers were detected by flow cytometry, immunofluorescence and western blotting.
RESULTS AND CONCLUSION: The adherent capacity of PP1-PP6 cells was gradually weakened after 6-step differential adhesion. After two to three days, PP6 cells began to be adherent and became round or spindle-shaped. Flow cytometry detected stem cell antigen 1 (+) 5.7%, embryonic antigen (+) 2.6%, and desmin (+) 41.2% in PP6 cells. Immunofluorescence exhibited that only a few cells expressed stem cell antigen 1 and embryonic antigen, showing diffused distribution. Stem cell antigen 1 was mainly expressed in cytoplasm, and embryonic antigen was mainly expressed in nuclei. Cell aggregation expressing desmin showed a large number and mainly expressed in cytoplasm. Very a few double-positive cells, such as stem cell antigen 1/embryonic antigen, stem cell antigen 1/desmin and embryonic antigen/desmin, were detected. No significant expression of stem cell antigen 1 and embryonic antigen protein was determined in PP1-PP5 cells. However, stem cell antigen 1/embryonic antigen protein was detected in PP6 cells. Desmin expression was gradually enhanced in PP1-PP6 cells. Results indicated that the cells from rat penile corpus cavernosum had the cell phenotype of stem cells and muscle-derived stem cells.