Anna Burdzińska1, Urszula Bartoszuk, Arkadiusz Orzechowski. 1. Department of Immunology, Transplantology, and Internal Disease, Transplantation Institute, Medical University of Warsaw, Warsaw, Nowogrodzka, Poland.
Abstract
OBJECTIVES: Myogenic cell therapy is considered a new method to treat urinary incontinence. One of the crucial limitations in cellular transplantation is poor cell survival after cell transfer. Preconditioning of cells before transfer is a new approach in improving the efficacy of graft. METHODS: In this study, autologous cell transplantation was performed on a rat model. Muscle-derived cells were isolated, identified with immunofluorescence and immunoblotting, preincubated or not with 10 ng/mL of basic fibroblast growth factor (bFGF) or 0.1 mM of sodium ascorbate for 24 hours, and transfected with the beta-galactosidase encoding gene using lipofectamine. Cell suspension was injected into the urethral wall. The tissue was collected 2 weeks after injection. RESULTS: The presence of transplanted cells was confirmed by X-Gal staining. Moreover, the effects of preincubation with either sodium ascorbate or bFGF 24 hours before transfer were evaluated. A semiqualitative analysis using chlorophenol-red-beta-d-galactopyranoside substrate demonstrated that preincubation with ascorbate had no effect on the number of transplanted cells but that bFGF significantly increased the amount of cells in the engrafted tissue by 46% (P < .05). CONCLUSIONS: Preconditioning of myogenic cells with bFGF should be seriously considered as a new tool for improving cell viability after cell transfer in the treatment of urinary incontinence.
OBJECTIVES: Myogenic cell therapy is considered a new method to treat urinary incontinence. One of the crucial limitations in cellular transplantation is poor cell survival after cell transfer. Preconditioning of cells before transfer is a new approach in improving the efficacy of graft. METHODS: In this study, autologous cell transplantation was performed on a rat model. Muscle-derived cells were isolated, identified with immunofluorescence and immunoblotting, preincubated or not with 10 ng/mL of basic fibroblast growth factor (bFGF) or 0.1 mM of sodium ascorbate for 24 hours, and transfected with the beta-galactosidase encoding gene using lipofectamine. Cell suspension was injected into the urethral wall. The tissue was collected 2 weeks after injection. RESULTS: The presence of transplanted cells was confirmed by X-Gal staining. Moreover, the effects of preincubation with either sodium ascorbate or bFGF 24 hours before transfer were evaluated. A semiqualitative analysis using chlorophenol-red-beta-d-galactopyranoside substrate demonstrated that preincubation with ascorbate had no effect on the number of transplanted cells but that bFGF significantly increased the amount of cells in the engrafted tissue by 46% (P < .05). CONCLUSIONS: Preconditioning of myogenic cells with bFGF should be seriously considered as a new tool for improving cell viability after cell transfer in the treatment of urinary incontinence.