The Fate of Implanted Syngenic Muscle Precursor Cells in Injured Striated Urethral Sphincter of Female Rats

Abstract

We studied the outcome of syngenic skeletal muscle precursor cells (MPCs) implanted in the striated urethral sphincter of the female rat. These cells were injected at the site of a longitudinal sphincterotomy performed 21 days before implantation. MPCs were isolated from the striated hindlimb muscles of syngenic adult rats and were infected with a retrovirus carrying the gene for either the green fluorescent protein (GFP) or the β-galactosidase enzyme (β-gal). MPCs (2 × 10⁵) were injected longitudinally at the site of the lesion in 48 animals using a 10-μl Hamilton syringe. Then the whole urethras were excised from 2 h up to 90 days for cross section immunocytochemistry analysis. All the urethras exhibited connective tissue in place of the injury of the striated fibers. Two hours after injection a cluster of small round basophilic cells was observable at the site of injection and some of them expressed GFP or β-gal. A few GFP- and β-gal-positive cells were already detectable 7 days after injection. A large amount of injected cells probably died after injection. Many striated fibers of the urethra became GFP positive from day 7 until day 21, suggesting that few MPCs were allowed to incorporate the divided extremities of the striated fibers from day 7. Unfortunately, we did not observe centronucleated regenerated fibers in this experiment.