RRC ID 82059
Author Yasuda M, Kato T, Okano M, Yamashita H, Matsuoka Y, Shirouzu Y, Fujioka T, Hattori F, Tsuji S, Kaneko K, Hitomi H.
Title Efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells, involving the inhibition of mTOR.
Journal Sci Rep
Abstract The mechanistic/mammalian target of rapamycin (mTOR) is involved in a wide range of cellular processes. However, the role of mTOR in podocytes remains unclear. In this study, we aimed to clarify the role of mTOR in podocyte differentiation from human induced pluripotent stem cells (hiPSCs) and to establish an efficient differentiation protocol for human podocytes. We generated podocytes from hiPSCs by modifying protocol. The expression of the podocyte-specific slit membrane components nephrin and podocin was measured using PCR, western blotting, flow cytometry, and immunostaining; and the role of mTOR was evaluated using inhibitors of the mTOR pathway. Nephrin and podocin were found to be expressed in cells differentiated from hiPSCs, and their expression was increased by mTOR inhibitor treatment. S6, a downstream component of the mTOR pathway, was also found to be involved in podocyte differentiation. we evaluated its permeability to albumin, urea, and electrolytes. The induced podocytes were permeable to the small molecules, but only poorly permeable to albumin. We have shown that the mTOR pathway is involved in podocyte differentiation. Our monolayer podocyte differential protocol, using an mTOR inhibitor, provides a novel in vitro model for studies of kidney physiology and pathology.
Volume 13(1)
Pages 20010
Published 2023-11-16
DOI 10.1038/s41598-023-47087-8
PII 10.1038/s41598-023-47087-8
PMID 37973990
PMC PMC10654390
MeSH Albumins / metabolism Cell Differentiation Humans Induced Pluripotent Stem Cells* / metabolism Kidney / metabolism MTOR Inhibitors Podocytes* / metabolism Sirolimus / pharmacology TOR Serine-Threonine Kinases / metabolism
Resource
Human and Animal Cells HiPS-RIKEN-2F(HPS0014)