Generation of human induced pluripotent stem cells from urine samples

Generation of human induced pluripotent stem cells from urine samples

Nature Protocols

PHOTO: Morphology of urinary cells at different time points after collection.
(a) Fresh female urine samples mainly consist of squamous cells and a few blood cells (arrowheads). (b) Fresh male urine samples mainly contain a few blood cells (arrowhead). (c–h) Type I (c,e,g) and type II (d,f,h) urinary cell colonies (arrowheads) at days 4 (c,d), 9 (e,f) and 12 (g,h). Representative images are shown in a–h; scale bars, 400 μm.

Human induced pluripotent stem cells (iPSCs) have been generated with varied efficiencies from multiple tissues. Yet, acquiring donor cells is, in most instances, an invasive procedure that requires laborious isolation. Here we present a detailed protocol for generating human iPSCs from exfoliated renal epithelial cells present in urine. This method is advantageous in many circumstances, as the isolation of urinary cells is simple (30 ml of urine are sufficient), cost-effective and universal (can be applied to any age, gender and race). Moreover, the entire procedure is reasonably quick—around 2 weeks for the urinary cell culture and 3–4 weeks for the reprogramming—and the yield of iPSC colonies is generally high—up to 4% using retroviral delivery of exogenous factors. Urinary iPSCs (UiPSCs) also show excellent differentiation potential, and thus represent a good choice for producing pluripotent cells from normal individuals or patients with genetic diseases, including those affecting the kidney.