Generation of human induced pluripotent stem cells from urine samples

Abstract

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.

Naïve Induced Pluripotent Stem Cells Generated From β-Thalassemia Fibroblasts Allow Efficient Gene Correction With CRISPR/Cas9

“…As demonstrated in previous reports, human urinary cells (hUCs) can be reprogrammed into primed iPSCs by introducing exogenous genes [24]; thus, we tested whether hUCs could be reprogrammed directly into the naïve iPSCs. Using a similar strategy, 5 3 10 5 hUCs obtained from a normal male were electroporated with the same episomal vectors and were then cultured in conventional hESM for 9 days.…”

“…Significant improvements in mutation correction efficiencies were achieved using a CRISPR/ Cas9 editing system in these naïve iPSCs, which are capable of hematopoietic differentiation. In addition, human naïve iPSCs could also be directly generated from noninvasively collected urinary cells [24], which are easily acquired and thus represent an excellent cell resource for further clinical trials. Thus, our study offers an improved strategy for personalized treatment of b-thalassemia and has important implications for the clinical use of human naïve iPSCs in the future.…”

Genetics of Human Sleep Behavioral Phenotypes

“…Recent advancement in stem cell research makes it possible to generate and propagate induced pluripotent stem cells (iPSCs) from skin biopsy or urine samples (containing exfoliated renal epithelial cells; Takahashi et al, 2007;Yu et al, 2007;Zhou et al, 2012). DNA can then be extracted from iPSC cultures for use in genetic analyses.…”

Induced Pluripotent Stem Cell Therapies for Degenerative Disease of the Outer Retina: Disease Modeling and Cell Replacement

“…iPSC have been so far reprogrammed from several different somatic cells types, including fibroblasts, 15,16 blood mononuclear cells, 17 peripheral blood cells, 18 human astrocytes, 19 renal epithelial cells present in urine, 20 and different types of cells associated with the hair (keratinocytes, 21 follicle dermal papilla, 22 and as reviewed in Ref. 23 ).…”