Tissue engineering of ureteral grafts by seeding urothelial differentiated hADSCs onto biodegradable ureteral scaffolds

Abstract

The study is aimed to evaluate the differentiation potential of human adipose-derived stem cells (hADSCs) into urothelial lineage, and to assess possibility of constructing ureteral grafts using the differentiated hADSCs and a novel polylactic acid (PLA)/collagen scaffolds. HADSCs were indirectly cocultured with urothelial cells in a transwell coculture system for urothelial differentiation. After 14 days coculturing, differentiation was evaluated by detecting urothelial lineage markers (cytokeratin-18 and uroplakin 2) in mRNA and protein level. Then the differentiated hADSCs were seeded onto PLA/collagen ureteral scaffolds and cultured in vitro for 1 week. The biocompatibility of the scaffolds was tested by scanning electron microscopy (SEM) and MTT analysis. At last, the cell/scafflod grafts were subcutaneously implanted into 4-week-old female athymic mice for 14 days. The results demonstrated that the hADSCs could be efficiently induced into urothelial lineage by indirect coculture. The differentiated cells seeded onto the PLA/collagen ureteral scaffolds survived up to 7 days and maintained proliferation in vitro, which indicated that the scaffolds displayed good biocompatibility. In vivo study showed that the differentiated cells in the grafts survived, formed multiple layers on the scaffolds and expressed urothelial lineage markers. In conclusion, hADSCs may serve as an alternative cell resource in cell-based tissue engineering for ureteral reconstruction. These cells could be employed to construct a model of ureteral engineering grafts and be effectively applied in vivo, which could be a new strategy on ureteral replacement with applicable potential in clinical research.

One-stage focal cartilage defect treatment with bone marrow mononuclear cells and chondrocytes leads to better macroscopic cartilage regeneration compared to microfracture in goats

“…The combination of cells derived from native tissue with other cell types has recently gained attention to address currently defined challenges in regenerative medicine 1 . For example, vascular endothelial cells and urothelial cells were both able to differentiate adipose-derived stem cells into respectively the osteogenic and urothelial lineage in order to enhance bone and urothelial tissue engineering 2,3 . Articular cartilage matrix production of expanded articular chondrocytes has been shown to be positively influenced by primary chondrocytes and human adipose-derived and embryonic stem cells in vitro 4e6 .…”

Isolation and Characterization of a Spontaneously Immortalized Multipotent Mesenchymal Cell Line Derived from Mouse Subcutaneous Adipose Tissue

“…For example, stem cells can now be used in cell therapy applications to obtain cell sheets or 3D proto-tissues under controlled conditions [24][25][26][27][28][29]. The latter are dictated by different local environment factors, such as growth-differentiation factors, adhesive molecules within the extracellular matrix, the properties of a possible supporting scaffold, as well as genetic programming and stochastic events [30][31][32][33][34][35][36][37][38][39][40]. All these studies may require a significant amount of primary cells and an extensive in vitro expansion, but ASCs generally undergo ''replicative senescence'' and irreversible growth arrest after a few replication cycles [20].…”

Ureteral reconstruction with reinforced collagen scaffolds in a porcine model

“…This way the initial tissue contraction takes place in a more controlled environment, without the toxic influences of urine (Adamowicz et al , ). Recent studies have shown the successful application of these techniques by implanting the tissue in non‐functional locations (Fu et al , ; Shi et al , ; Xu et al , ), as well as using them for ureteral repair afterwards (Liao et al , ; Zhang et al , ). Pre‐implantation before functional implantation may therefore improve the outcome of the reconstruction.…”