Efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells, involving the inhibition of mTOR
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)...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-11-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-47087-8 |
| _version_ | 1827711432050868224 |
|---|---|
| author | Masahiro Yasuda Tadashi Kato Mai Okano Hiromi Yamashita Yoshikazu Matsuoka Yasumasa Shirouzu Tatsuya Fujioka Fumiyuki Hattori Shoji Tsuji Kazunari Kaneko Hirofumi Hitomi |
| author_facet | Masahiro Yasuda Tadashi Kato Mai Okano Hiromi Yamashita Yoshikazu Matsuoka Yasumasa Shirouzu Tatsuya Fujioka Fumiyuki Hattori Shoji Tsuji Kazunari Kaneko Hirofumi Hitomi |
| author_sort | Masahiro Yasuda |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-10T17:55:35Z |
| format | Article |
| id | doaj.art-1214bdda34164203b7da890f5b895cce |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-10T17:55:35Z |
| publishDate | 2023-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-1214bdda34164203b7da890f5b895cce2023-11-20T09:12:32ZengNature PortfolioScientific Reports2045-23222023-11-0113111210.1038/s41598-023-47087-8Efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells, involving the inhibition of mTORMasahiro Yasuda0Tadashi Kato1Mai Okano2Hiromi Yamashita3Yoshikazu Matsuoka4Yasumasa Shirouzu5Tatsuya Fujioka6Fumiyuki Hattori7Shoji Tsuji8Kazunari Kaneko9Hirofumi Hitomi10Department of iPS Stem Cell Regenerative Medicine, Kansai Medical UniversityDepartment of iPS Stem Cell Regenerative Medicine, Kansai Medical UniversityDepartment of iPS Stem Cell Regenerative Medicine, Kansai Medical UniversityDepartment of iPS Stem Cell Regenerative Medicine, Kansai Medical UniversityDepartment of iPS Stem Cell Regenerative Medicine, Kansai Medical UniversityDepartment of iPS Stem Cell Regenerative Medicine, Kansai Medical UniversityDepartment of iPS Stem Cell Regenerative Medicine, Kansai Medical UniversityDepartment of iPS Stem Cell Regenerative Medicine, Kansai Medical UniversityDepartment of Pediatrics, Kansai Medical UniversityDepartment of Pediatrics, Kansai Medical UniversityDepartment of iPS Stem Cell Regenerative Medicine, Kansai Medical UniversityAbstract 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.https://doi.org/10.1038/s41598-023-47087-8 |
| spellingShingle | Masahiro Yasuda Tadashi Kato Mai Okano Hiromi Yamashita Yoshikazu Matsuoka Yasumasa Shirouzu Tatsuya Fujioka Fumiyuki Hattori Shoji Tsuji Kazunari Kaneko Hirofumi Hitomi Efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells, involving the inhibition of mTOR Scientific Reports |
| title | Efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells, involving the inhibition of mTOR |
| title_full | Efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells, involving the inhibition of mTOR |
| title_fullStr | Efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells, involving the inhibition of mTOR |
| title_full_unstemmed | Efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells, involving the inhibition of mTOR |
| title_short | Efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells, involving the inhibition of mTOR |
| title_sort | efficient protocol for the differentiation of kidney podocytes from induced pluripotent stem cells involving the inhibition of mtor |
| url | https://doi.org/10.1038/s41598-023-47087-8 |
| work_keys_str_mv | AT masahiroyasuda efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor AT tadashikato efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor AT maiokano efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor AT hiromiyamashita efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor AT yoshikazumatsuoka efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor AT yasumasashirouzu efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor AT tatsuyafujioka efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor AT fumiyukihattori efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor AT shojitsuji efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor AT kazunarikaneko efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor AT hirofumihitomi efficientprotocolforthedifferentiationofkidneypodocytesfrominducedpluripotentstemcellsinvolvingtheinhibitionofmtor |