The goal of this study was to determine whether urothelial cells (UC) and smooth muscle cells (SMC) derived from the differentiation of urine-derived stem cells (USC) could be used to form engineered urethral tissue when seeded on a modified 3-D porous small intestinal submucosa (SIS) scaffold. Cells were obtained from 12 voided urine samples from 4 healthy individuals. USC were isolated, characterized and induced to differentiate into UC and SMC. Fresh SIS derived from pigs was decellularized with 5% peracetic acid (PAA). Differentiated UC and SMC derived from USC were seeded onto SIS scaffolds with highly porous microstructure in a layered co-culture fashion and cultured under dynamic conditions for one week. The seeded cells formed multiple uniform layers on the SIS and penetrated deeper into the porous matrix during dynamic culture. USC that were induced to differentiate also expressed UC markers (Uroplakin-III and AE1/AE3) or SMC markers (α-SM actin, desmin, and myosin) after implantation into athymic mice for one month, and the resulting tissues were similar to those formed when UC and SMC derived from native ureter were used. In conclusion, UC and SMC derived from USC could be maintained on 3-D porous SIS scaffold. The dynamic culture system promoted 3-D cell-matrix ingrowth and development of a multilayer mucosal structure similar to that of native urinary tract tissue. USC may serve as an alternative cell source in cell-based tissue engineering for urethral reconstruction or other urological tissue repair.
"Cells differentiated from USCs were seeded on a modified 3D porous small intestinal submucosa in order to form engineered urethral tissue. Results showed that the cells formed multiple uniform layers on the scaffolds, which was similar to that of native urinary tract tissue . As USCs can be obtained using non-invasive and simple methods, they represent a promising alternative stem cell population for tissue regeneration. "
"These cells were able to differentiate into smooth muscle and urothelial cells in vitro, and after seeding on bacterial cellulose scaffold, were used to create model of urinary conduit . One year later, this same method, using 3-D porous SIS scaffold , was used for experimental urethra construction . Due to its ability to differentiate into urothelial lineages, USCs are an ideal cell source for the regeneration of other urinary tract components like the bladder, or the ureters  . "
"Urine-derived stem cells (USCs) are first described by Zhang et al,3 and are thought to derive from basal cells of the urothelial layer. They can be isolated from urine and expanded in special culture, can differentiate into cell lineages such as urothelial cells, smooth muscle cells, endothelial cells, osteocytes, chondrocytes, and adipocytes on induction with appropriate media,4–8 and have been reported recently to be ideal candidates for mesenchymal stem cells to replace marrow-derived stem cells and adipose-derived stem cells as seed cells for tissue engineering, due to their harvest from autologous urine using noninvasive, safe, low-cost, and easily reproducible methods.3,7 Currently, USCs are being investigated as cell therapies in the treatment of renal insufficiency and urinary incontinence, and for tissue engineering in bladder and urethral tissue regeneration;6 however, their use in tissue-engineering to repair bone defects is equally attractive. "
[Show abstract][Hide abstract] ABSTRACT: In tissue engineering, urine-derived stem cells are ideal seed cells and silver nanoparticles (AgNPs) are perfect antimicrobial agents. Due to a distinct lack of information on the effects of AgNPs on urine-derived stem cells, a study was conducted to evaluate the effects of silver ions and AgNPs upon the cytotoxicity and osteogenic differentiation of urine-derived stem cells. Initially, AgNPs or AgNO3 were exposed to urine-derived stem cells for 24 hours. Cytotoxicity was measured using the Cell Counting kit-8 (CCK-8) test. The effects of AgNPs or AgNO3 at the maximum safety concentration determined by the CCK-8 test on osteogenic differentiation of urine-derived stem cells were assessed by alkaline phosphatase activity, Alizarin Red S staining, and the quantitative reverse transcription polymerase chain reaction. Lastly, the effects of AgNPs or AgNO3 on "urine-derived stem cell actin cytoskeleton organization" and RhoA activity were assessed by rhodamine-phalloidin staining and Western blotting. Concentration-dependent toxicity was observed starting at an AgNO3 concentration of 2 μg/mL and at an AgNP concentration of 4 μg/mL. At these concentrations, AgNPs were observed to promote osteogenic differentiation of urine-derived stem cells, induce actin polymerization and increase cytoskeletal tension, and activate RhoA; AgNO3 had no such effects. In conclusion, AgNPs can promote osteogenic differentiation of urine-derived stem cells at a suitable concentration, independently of silver ions, and are suitable for incorporation into tissue-engineered scaffolds that utilize urine-derived stem cells as seed cells.
International Journal of Nanomedicine 05/2014; 9(1):2469-2478. DOI:10.2147/IJN.S59753 · 4.38 Impact Factor
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