Biocompatibility of Wharton's jelly mesenchymal stem cells with porcine bladder accellular matrix

Objective To explore the biocompatibility of porcine bladder accellular matrix (PBAM) prepared by chemical method and human umbilical cord Wharton mesenchymal stem cells (WJ-MSCs), and provide new seed cells for further research of tissueengineered bladder. Methods PBAM was prepared by chemical decellular method, and HE, toluidine blue and Hoechst33258 fluorescent staining were used to detect the effect of the decellularization. The appearance of PBAM was observed by scanning electron microscope and the porosity was detected by anhydrous ethanol. WJ-MSCs were cultured by tissue block method, and its morphology was observed, then osteogenic and adipogenic differentiation were induced. The effects of different concentrations of PBAM leaching liquor on the proliferation of WJ-MSCs were compared. The experimental group combined WJ-MSCs to PBAM for culture, and the control group was cultured by the DMEM. At 3 days, 5 days, 7 days of cultivation, the WJ-MSCs were collected and counted. The cell-scaffold was observed by Hoechst33258 and AO staining. And the growing of WJ-MSCs on PBAM was observed by scanning electron microscope (SEM). Results PBAM was successfully prepared by chemical acellular method. HE, toluidine blue and Hoechst33258 staining confirmed no cell residue on PBAM. SEM showed a porous structure on the surface of PBAM, and the porosity of PBAM was 93%. At 6 days of cultivation, the WJ-MSCs were fibroblast-like, and after osteogenic and adipogenic induction, the results of oil red “O” and Alizarin Red stain were positive. The effect of different concentrations of PBAM leaching liquor on the proliferation of WJ-MSCs was similar to that of DMEM. The proliferation peaked at the fifth day in both two groups, and the proliferation rate of WJ-MSCs in the experimental group was higher than that of the control group (P＜ 0.05). At the fifth day after WJ-MSCs combined to the PBAM, Hoechst33258 staining showed that a large number of WJ-MSCs grew on the surface of PBAM, and AO staining suggested that they were living cells. SEM showed that a large number of WJ-MSCs adhered and grown. Conclusion PBAM shows good compatibility with WJ-MSCs, which provides an experimental basis for further study of tissue-engineered bladder.