Differentiation Of Urine-derived Stem Cells Research Articles

Renal clear cancer cells regulate the differentiation of urine-derived stem cells to carcinoma-associated fibroblasts via RUNX3/TGF-β1 signaling axis.

Clear cell renal cell carcinoma (ccRCC), the most common pathological subtype of renal cancer, is one of the significant health concerns due to limited clinically effective treatments. Nevertheless, targeting carcinoma-associated fibroblasts in the tumor microenvironment has emerged as a promising innovative strategy for renal cancer therapy. Thus, this study is aimed to explore the role and molecular mechanism of urine-derived stem cells (USCs) in the progression and metastasis of ccRCC. Initially, wound-healing and transwell experiments were used to assess the migration and invasion abilities of the cells. Then, western blot analysis (WB) and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analyses were used to demonstrate the relevant protein and messenger RNA expression levels. Finally, hematoxylin-eosin and immunohistochemical stainings were performed to evaluate metastasis and protein expression in lung tumors. The coculture of USCs with the ccRCC cell lines significantly enhanced their migratory and invasive abilities. WB and qRT-PCR analyses exhibited that ccRCC cell lines significantly increased cell mobility markers transcriptional and protein levels in USCs. Finally, the in vivo investigations in nude mice showed that USCs promoted the proliferation and migration of ccRCC-based xenograft tumors. In summary, these findings demonstrated that USCs promoted ccRCC tumorigenesis and development in vivo and in vitro by regulating the Runt-related transcription factor 3/transforming growth factor-β1 signaling axis.

Urine-Derived Stem Cells for Epithelial Tissues Reconstruction and Wound Healing.

Epithelial tissue injury can occur on any surface site of the body, particularly in the skin or urethral mucosa tissue, due to trauma, infection, inflammation, and toxic compounds. Both internal and external body epithelial tissue injuries can significantly affect patients’ quality of life, increase healthcare spending, and increase the global economic burden. Transplantation of epithelial tissue grafts is an effective treatment strategy in clinical settings. Autologous bio-engineered epithelia are common clinical skin substitutes that have the specific advantages of avoiding tissue rejection, obviating ethical concerns, reducing the risk of infection, and decreasing scarring compared to donor grafts. However, epithelial cells are often obtained from the individual’s skin and mucosa through invasive methods, which cause further injury or damage. Urine-derived stem cells (USC) of kidney origin, obtained via non-invasive acquisition, possess high stemness properties, self-renewal ability, trophic effects, multipotent differentiation potential, and immunomodulatory ability. These cells show versatile potential for tissue regeneration, with extensive evidence supporting their use in the repair of epidermal and urothelial injuries. We discuss the collection, isolation, culture, characterization, and differentiation of USC. We also discuss the use of USC for cellular therapies as well as the administration of USC-derived paracrine factors for epidermal and urothelial tissue repair. Specifically, we will discuss 3D constructions involving multiple types of USC-loaded hydrogels and USC-seeded scaffolds for use in cosmetic production testing, drug development, and disease modeling. In conclusion, urine-derived stem cells are a readily accessible autologous stem cell source well-suited for developing personalized medical treatments in epithelial tissue regeneration and drug testing.

Nanotopographical 3D-Printed Poly(ε-caprolactone) Scaffolds Enhance Proliferation and Osteogenic Differentiation of Urine-Derived Stem Cells for Bone Regeneration.

3D-printing technology can be used to construct personalized bone substitutes with customized shapes, but it cannot regulate the topological morphology of the scaffold surface, which plays a vital role in regulating the biological behaviors of stem cells. In addition, stem cells are able to sense the topographical and mechanical cues of surface of scaffolds by mechanosensing and mechanotransduction. In our study, we fabricated a 3D-printed poly(ε-caprolactone) (PCL) scaffold with a nanotopographical surface and loaded it with urine-derived stem cells (USCs) for application of bone regeneration. The topological 3D-printed PCL scaffolds (TPS) fabricated by surface epiphytic crystallization, possessed uniformly patterned nanoridges, of which the element composition and functional groups of nanoridges were the same as PCL. Compared with bare 3D-printed PCL scaffolds (BPS), TPS have a higher ability for protein adsorption and mineralization in vitro. The proliferation, cell length, and osteogenic gene expression of USCs on the surface of TPS were significantly higher than that of BPS. In addition, the TPS loaded with USCs exhibited a good ability for bone regeneration in cranial bone defects. Our study demonstrated that nanotopographical 3D-printed scaffolds loaded with USCs are a safe and effective therapeutic strategy for bone regeneration.

Silver nanoparticles promote osteogenic differentiation of human urine-derived stem cells at noncytotoxic concentrations.

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.

Skeletal myogenic differentiation of urine-derived stem cells and angiogenesis using microbeads loaded with growth factors

To provide site-specific delivery and targeted release of growth factors to implanted urine-derived stem cells (USCs), we prepared microbeads of alginate containing growth factors. The growth factors included VEGF, IGF-1, FGF-1, PDGF, HGF and NGF. Radiolabeled growth factors were loaded separately and used to access the in vitro release from the microbeads with a gamma counter over 4 weeks. In vitro endothelial differentiation of USCs by the released VEGF from the microbeads in a separate experiment confirmed that the released growth factors from the microbeads were bioactive. USCs and microbeads were mixed with the collagen gel type 1 (2 mg/ml) and used for in vivo studies through subcutaneous injection into nude mice. Four weeks after subcutaneous injection, we found that grafted cell survival was improved and more cells expressed myogenic and endothelial cell transcripts and markers compared to controls. More vessel formation and innervations were observed in USCs combined with six growth factors cocktail incorporated in microbeads compared to controls. In conclusion, a combination of growth factors released locally from the alginate microbeads induced USCs to differentiate into a myogenic lineage, enhanced revascularization and innervation, and stimulated resident cell growth in vivo. This approach could potentially be used for cell therapy in the treatment of stress urinary incontinence.

1609 POTENTIAL USE FOR UROTHELIAL MUCOSA ENGINEERED WITH AUTOLOGOUS URINE DERIVED STEM CELLS SEEDED ON A POROUS COLLAGEN MATRIX IN HYPOSPADIAS REPAIR

You have accessJournal of UrologyPediatrics: Congenital Anomalies - Lower Urinary Tract & Genitalia II1 Apr 20121609 POTENTIAL USE FOR UROTHELIAL MUCOSA ENGINEERED WITH AUTOLOGOUS URINE DERIVED STEM CELLS SEEDED ON A POROUS COLLAGEN MATRIX IN HYPOSPADIAS REPAIR Rongpei Wu, Guihua Liu, Shaofeng Wu, Anthony Atala, and Yuanyuan Zhang Rongpei WuRongpei Wu Winston Salem, NC More articles by this author , Guihua LiuGuihua Liu Winston Salem, NC More articles by this author , Shaofeng WuShaofeng Wu Winston Salem, NC More articles by this author , Anthony AtalaAnthony Atala Winston Salem, NC More articles by this author , and Yuanyuan ZhangYuanyuan Zhang Winston Salem, NC More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1404AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Surgical repair of hypospadias often requires autogenous tissue, such as foreskin, bladder or oral mucosa to extend urethral segments. However, the procedure required to obtain these tissues is an invasive approach and it may lead to donor site morbidity. Additionally, adequate tissue for this substitution is not always available, and if the urethral defect is too long, this type of repair is not possible. Tissue-engineered tubular urethral tissues are an alternative option and allow for replacement of deficient urethral tissues with functionally equivalent ones. The goal of this study was to generate a 3-dimensional porous small intestinal submucosa (SIS) scaffold and recellularize this scaffold with urothelial cells (UC) and smooth muscle cells (SMC) derived from the differentiation of urine-derived stem cells (USC) in order to form engineered urethral tissue that could be used in hypospadias repair. METHODS Fresh SIS derived from pigs was decellularized with 5% peracetic acid (PAA). Cells were obtained from voided urine samples from healthy individuals. USC were isolated, characterized and induced to differentiate into UC and SMC. Differentiated UC and SMC derived from USC were seeded onto the SIS scaffolds 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. Urothelial and muscle cells derived from USC expressed UC markers (Uroplakin-III, AE1/AE3) or SMC markers (α-SM actin, desmin, and myosin), respectively, after implantation into athymic mice. RESULTS DNA content analysis showed that most nuclear material was removed from the SIS scaffold. Treatment with 5% PAA led to development of a highly porous surface with retention of less cellular material, and the decellularized SIS maintained the majority of the tensile strength of normal tissue. In 3-D dynamic culture, the seeded cells (UC and SMC) formed multiple, even layers on the surface of matrix. Cells also penetrated deeper into the lamina propria of the matrix compared to untreated matrix. Immunocytochemical staining indicated that the grafted USC expressed urothelial- and smooth muscle-specific markers both, in vitro and in vivo. CONCLUSIONS This study demonstrates that the decellularized SIS possesses appropriate 3-D porosity to prompt the induced USC to form multilayer of urothelium and SMC-matrix infiltration, which provides potential applications in urethral reconstruction in hypospdias repair. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e651 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Rongpei Wu Winston Salem, NC More articles by this author Guihua Liu Winston Salem, NC More articles by this author Shaofeng Wu Winston Salem, NC More articles by this author Anthony Atala Winston Salem, NC More articles by this author Yuanyuan Zhang Winston Salem, NC More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Human urine-derived stem cells seeded in a modified 3D porous small intestinal submucosa scaffold for urethral tissue engineering

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.