Footprint- and xeno-free human iPSCs derived from urine cells using extracellular matrix-based culture conditions

Abstract

The efficient generation of integration- and xeno-free iPSCs is a prerequisite for their use in clinical applications. Furthermore, non-invasiveness of somatic cell acquisition for iPSC generation is another factor to consider. In this study, we established a practical, simple, and convenient method to generate integration- and xeno-free iPSCs from urine cells which can be obtained in a non-invasive manner. Our method was based on extracellular matrix-based xeno-free iPSC culture condition and episomal transfection, and worked efficiently with both urine cells and adipose-derived stromal cells (ADSCs). To obtain strictly xeno-free iPSCs, we also formulated a new xeno-free culture medium for primary urine cells. Intriguingly, urine cells displayed slower growth, and more dramatic increase in apoptosis at high passage numbers than ADSCs. However, urine cells at low passage (<P3) displayed modest apoptosis (∼7–8%) and relatively high (∼0.29%) efficiency of iPSC generation.This study reports the generation of integration- and xeno-free iPSCs from non-invasively obtained urine cells.