Isolation and Characterization of Human Muscle-derived Cells

Objectives: To isolate and characterize human muscle-derived cells (MDCs) for future management applications on lower urinary tract symptoms, including stress urinary incontinence and bladder reconstitution. The development of muscle stem cells for transplantation or gene transfer in patients with muscle disorders has become more attractive and challenging recently. Methods: Human MDCs were isolated from the skeletal muscles of the limbs. The muscle tissues were minced, digested at 37°C by 0.2% collagenase, trypsinized, filtered, and cultured in F12 medium with 15% fetal bovine serum at 37°C. Human MDCs were then isolated using a modified preplate technique. After isolation, the MDCs were characterized by immunohistochemistry, flow cytometry, and indirect immunofluorescence. Results: The growth doubling time of the MDCs was approximately 24 hours. Immunohistochemistry study was performed with the stem cell markers CD34, CD117, vascular cell adhesion molecule, and vascular endothelial growth factor receptor 2, and the relative stem cell position was identified. Positive immunofluorescence outcomes were found with the stem cell markers, myoblast markers CXCR4, CD56, desmin, and a fibroblast marker AB-1. Flow cytometry analysis identified markers CD34 and CD56 in the isolated MDCs, with a percentage of 5.12% and 10.34%, respectively. Conclusions: The isolation and characterization of human MDCs was successfully achieved. Human MDCs might have the potential to be a novel tool for the management of stress urinary incontinence and bladder reconstitution.