Generation of infant- and pediatric-derived urinary induced pluripotent stem cells competent to form kidney organoids

• Published in: Pediatric research Vol. 87; no. 4; pp. 647 - 655
• Main Authors: Mulder, Jaap, Sharmin, Sazia, Chow, Theresa, Rodrigues, Deivid Carvalho, Hildebrandt, Matthew R, D'Cruz, Robert, Rogers, Ian, Ellis, James, Rosenblum, Norman D
• Format: Journal Article
• Language: English
• Published: United States Nature Publishing Group 01.03.2020
• Subjects: Adolescent; Case-Control Studies; Cell Proliferation; Cell Separation; Cells, Cultured; Cellular Reprogramming; Cellular Reprogramming Techniques; Child; Child, Preschool; Feasibility Studies; Female; Gene Expression Regulation, Developmental; Humans; Induced Pluripotent Stem Cells - metabolism; Induced Pluripotent Stem Cells - pathology; Infant; Kidney - metabolism; Kidney - pathology; Male; Organoids - metabolism; Organoids - pathology; Pediatrics; Phenotype; Stem cells; Urine; Urine - cytology; Urogenital Abnormalities - genetics; Urogenital Abnormalities - metabolism; Urogenital Abnormalities - pathology; Vesico-Ureteral Reflux - genetics; Vesico-Ureteral Reflux - metabolism; Vesico-Ureteral Reflux - pathology
• ISSN: 0031-3998; 1530-0447
• DOI: 10.1038/s41390-019-0618-y

Human induced pluripotent stem cells (iPSCs) are a promising tool to investigate pathogenic mechanisms underlying human genetic conditions, such as congenital anomalies of the kidney and urinary tract (CAKUT). Currently, iPSC-based research in pediatrics is limited by the invasiveness of cell collection. Urine cells (UCs) were isolated from pediatric urine specimens, including bag collections, and reprogrammed using episomal vectors into urinary iPSCs (UiPSCs). Following iPSC-quality assessment, human kidney organoids were generated. UCs were isolated from 71% (12/17) of single, remnant urine samples obtained in an outpatient setting (patients 1 month-17 years, volumes 10-75 ml). Three independent UCs were reprogrammed to UiPSCs with early episome loss, confirmed pluripotency and normal karyotyping. Subsequently, these UiPSCs were successfully differentiated into kidney organoids, closely resembling organoids generated from control fibroblast-derived iPSCs. Importantly, under research conditions with immediate sample processing, UC isolation was successful 100% for target pediatric CAKUT patients and controls (11/11) after at most two urine collections. Urine in small volumes or collected in bags is a reliable source for reprogrammable somatic cells that can be utilized to generate kidney organoids. This constitutes an attractive approach for patient-specific iPSC research involving infants and children with wide applicability and a low threshold for participation.