Generation of Three-Dimensional Renal Micro-Tis-sue with Co-Culture of Human Pluripotent Stem Cell-De-rived Renal Precursors and Supportive Cells

Objective: Directed differentiation of human pluripotent stem cell (hPSC) using a growing number of small molecules and growth factors is required for in vitro generation of renal line-age cells. By combining two-dimensional (۲D) protocol with a three-dimensional (۳D) co-culture system, we established a low-cost and rapid method for generation of a ۳D renal model from hPSCs.Materials and Methods: To determine whether a cost-effec-tive small molecule, inducer of definitive endoderm ۱ (IDE۱), can be used to replace activin A, we examined gene expression of multiple renal lineage markers in cells differentiated with IDE۱. In the following, we employed a ۳D co-culture system in which hPSC-derived kidney precursors were co-cultured with endothelial cells and mesenchymal stem cells (named RMEM). hPSC-derived kidney precursors were cultured either alone (re-nal micro-tissue (RM)) or in co-culture with Human umbili-cal vein endothelial cells (HUVECs) and bone marrow-derived mesenchymal stem cells (BM-MSCs) for ۱۱ days.Results: Data showed the expression of nephrogenic markers was significantly increased in cells differentiated with IDE۱ compared with cells differentiated using activin A. These data indicated that renal lineage cells can indeed be generated using this alternative approach. Immunofluorescent staining showed the expression of kidney specific markers, as well as endothe-lial cells that distributed throughout RMEMs. Quantitative Real-Time PCR analysis confirmed a significant increase in gene expression of some renal-specific markers in RMEMs. Conclusion: These findings demonstrated that renal precursors in co-culture with endothelial cells and mesenchymal stem cells showed greater maturity compared with micro-tissue without these cells.

کلیدواژه ها:

Renal Micro-Tissue ، Endothelial Cell ، Human Pluri-potent Stem Cells ، Differentiation ، Mesenchymal Stem Cell