Postdoc Seoul National University Hospital Center for Medical Research and Innovation, Seoul-t'ukpyolsi, Republic of Korea
Abstract: Inadequate knowledge of the fundamental mechanisms underlying pediatric neurological disorders impedes their effective treatment. Induced pluripotent stem cells (iPSCs) are essential to explore the course of neurological diseases because they enable the modeling of these diseases at the cellular level. This study aimed to generate an iPSC bank from urine cells (UCs) for clinical applications, especially for the study of pediatric neurogenetic diseases. Urine sample collection can benefit the large donor population because it is a noninvasive, painless, and simple technique, providing a plentiful cell source for iPSC generation. UCs were isolated from the urine of donors with specific diseases (n = 12; 7 males and 5 females). UCs were reprogrammed into iPSCs using episomal plasmid vectors and key transcription factors (OCT3/4, SOX2, KLF4, L-MYC, and LIN28). qPCR and immunocytochemistry confirmed the expression of pluripotent genes (OCT3/4, SOX2, NANOG, and LIN28) and pluripotent proteins (OCT4, NANOG, SSEA-4, and TRA-1-60). Trilineage differentiation potential was demonstrated by immunostaining embryonic-body-derived iPSCs for β-tubulin III, smooth muscle actin (SMA), and alpha-fetoprotein (AFP). Chromosomal microarray (CMA) was used to assess genomic stability, revealing pathogenic chromosomal deletions or duplications in four out of 12 lines. Notably, repeated CMA testing on earlier-passage lines showed normal genomic profiles in one of the affected lines, emphasizing the importance of genetic screening at multiple stages during iPSC culturing. This study successfully generated an iPSC bank derived from UCs of patients with early-onset neurogenetic diseases. The bank provides a robust, efficient protocol to expand access to patient-specific iPSCs, facilitating pediatric neurogenetic research and enabling disease modeling for the development of targeted therapies.
Funding Source: This work was funded by the New Faculty Startup Fund from Seoul National University and SNUH Lee Kun-Hee Child Cancer & Rare Disease Project, Republic of Korea (grant number: 24C017-0100).
