Improvement of transfection with reprogramming factors in urine-derived cells

Abstract

Human-induced pluripotent stem cells (iPSCs) are an accessible source of adult-derived, patient-specific pluripotent stem cells for use in basic research, drug discovery, disease modeling, and stem cell therapy. Improving the accessibility of methods to obtain iPSCs regardless of the cell source can enhance their clinical application. Therefore, our purpose is to report a simple protocol to obtain iPS-like cells from urine-derived renal epithelial cells (RECs) using different extracellular matrices and transfection reagents. In this study, we began by culturing urine-derived cells from healthy donors to establish a primary culture of renal epithelial cells, followed by their characterization. Subsequently, we generated iPS-like cells by transfecting renal epithelial cells (RECs) with vectors expressing Oct4, Sox2, L-Myc, Lin-28, and Klf4, and we compared the efficacy of different extracellular matrices and transfection reagents. The resultant iPS-like cells showed a human embryonic stem cell-like morphology and expressed the specific pluripotency markers Oct3/4, Nanog, Lin28, and Klf4. We concluded that Lipofectamine Stem Cell transfection reagent is more effective than FuGENE in obtaining iPS-like cells under the conditions tested. Moreover, the three matrices are similar in their efficiency of obtaining iPS-like cells. This report provides an experimental protocol for obtaining and generating iPS-like cells from urine samples for further cell therapy research on different human diseases.