The Experimental Research On Ureter Replacement Using BAM With ADSCs Seeding

Objective To investigate the clinical possibility and effect of substitution of ureter using ADSCs-seeding bladder acellular matrix and find an ideal replacement material to repair ureteral defect.Methods The first section:The subcutaneous adipose tissue was collected from the operation. Primary hADSCs were isolated subsequently by the methods of stirring digestion within1%collagenase type â… . The cells were divided into three groups and cultured in high glucose DMEM, high glucose DMEM with basic fibroblast growth factor (bFGF) and mesenchymal stem cell media (MSCM) respectively. The expressions of CD31, CD34and Stro-1were detected by immumofluorescence and flow cytometry method. The growth state of cells in the three groups was evaluated. Proliferative speed of cells in each group was evaluated after cultured in different culture conditions. The second section:The ADSCs of the porcine were seeded on BAM and cultured for about3days.6porcine were divided into2groups randomly and marked with A and B. The defects of group A were repaired with BAM, group B were repaired using ADSCs seeded BAM. IVP was performed in the third month postoperatively. Renal function and patency of new ureter were evlulated. After the IVP, all the porcine were killed, and the replaced ureter were resected through abdominal incision and then examined with pathologic method and immunohistochemistry.Results hADSCs were successfully isolated through stirring digestion within collagenase. The results of immunofluorescence and flow cytometry indicated that hADSCs of three groups were positive for Stro-1and CD34except CD31. Number of hADSCs that cultured in MSCM and high glucose DMEM with bFGF was more and the cell growth was significantly better than those cultured in high glucose DMEM (p<0.01). MSCM and high glucose DMEM with bFGF were significantly improve the proliferation of cells in each group than high glucose DMEM (p<0.05). None of the porcine died after operation in6cases of ureteral replacement. Portion of BAM was absorbed after3months, but the BAM was almost absorbed in group B. The uroepithelial cells, musculature and new blood vessels could be observed in group B, but little in group A. The lumen of replaced part was obstruction, and no contrast media circulated on IVP in group A. In group B, the IVP showed a stenotic ureteral lumen and delayed excretion with wild hydroureteronephrosis.Conclusion A great number of hADSCs could be obtained in a short time when cultured in MSCM and high glucose DMEM with bFGF, which can suffice for the cell source for tissue engineering. The ADSCs-seeded BAM could keep the channel function without clear contracture and necrosis, and showed a normal ureteric mucous and muscular layer while it was used to reconstruct serious ureteric defect. Therefore, the ADSCs-seeded BAM appeared the potential valve of being used as the scaffold in urology tissue engineering.