Generation of functional podocytes from human induced pluripotent stem cells
Generating human podocytes in vitro could offer a unique opportunity to study human diseases. Here, we describe a simple and efficient protocol for obtaining functional podocytes in vitro from human induced pluripotent stem cells. Cells were exposed to a three-step protocol, which induced their diff...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2016-07-01
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| Series: | Stem Cell Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506116300642 |
| _version_ | 1818138630849298432 |
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| author | Osele Ciampi Roberto Iacone Lorena Longaretti Valentina Benedetti Martin Graf Maria Chiara Magnone Christoph Patsch Christodoulos Xinaris Giuseppe Remuzzi Ariela Benigni Susanna Tomasoni |
| author_facet | Osele Ciampi Roberto Iacone Lorena Longaretti Valentina Benedetti Martin Graf Maria Chiara Magnone Christoph Patsch Christodoulos Xinaris Giuseppe Remuzzi Ariela Benigni Susanna Tomasoni |
| author_sort | Osele Ciampi |
| collection | DOAJ |
| description | Generating human podocytes in vitro could offer a unique opportunity to study human diseases. Here, we describe a simple and efficient protocol for obtaining functional podocytes in vitro from human induced pluripotent stem cells. Cells were exposed to a three-step protocol, which induced their differentiation into intermediate mesoderm, then into nephron progenitors and, finally, into mature podocytes. After differentiation, cells expressed the main podocyte markers, such as synaptopodin, WT1, α-Actinin-4, P-cadherin and nephrin at the protein and mRNA level, and showed the low proliferation rate typical of mature podocytes. Exposure to Angiotensin II significantly decreased the expression of podocyte genes and cells underwent cytoskeleton rearrangement. Cells were able to internalize albumin and self-assembled into chimeric 3D structures in combination with dissociated embryonic mouse kidney cells. Overall, these findings demonstrate the establishment of a robust protocol that, mimicking developmental stages, makes it possible to derive functional podocytes in vitro. |
| first_indexed | 2024-12-11T10:15:15Z |
| format | Article |
| id | doaj.art-167c2c0e6090410e8c10f32e85ce8309 |
| institution | Directory Open Access Journal |
| issn | 1873-5061 1876-7753 |
| language | English |
| last_indexed | 2024-12-11T10:15:15Z |
| publishDate | 2016-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj.art-167c2c0e6090410e8c10f32e85ce83092022-12-22T01:11:37ZengElsevierStem Cell Research1873-50611876-77532016-07-0117113013910.1016/j.scr.2016.06.001Generation of functional podocytes from human induced pluripotent stem cellsOsele Ciampi0Roberto Iacone1Lorena Longaretti2Valentina Benedetti3Martin Graf4Maria Chiara Magnone5Christoph Patsch6Christodoulos Xinaris7Giuseppe Remuzzi8Ariela Benigni9Susanna Tomasoni10IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, ItalyRoche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandIRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, ItalyIRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, ItalyRoche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandIRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, ItalyIRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, ItalyIRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, ItalyIRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, ItalyGenerating human podocytes in vitro could offer a unique opportunity to study human diseases. Here, we describe a simple and efficient protocol for obtaining functional podocytes in vitro from human induced pluripotent stem cells. Cells were exposed to a three-step protocol, which induced their differentiation into intermediate mesoderm, then into nephron progenitors and, finally, into mature podocytes. After differentiation, cells expressed the main podocyte markers, such as synaptopodin, WT1, α-Actinin-4, P-cadherin and nephrin at the protein and mRNA level, and showed the low proliferation rate typical of mature podocytes. Exposure to Angiotensin II significantly decreased the expression of podocyte genes and cells underwent cytoskeleton rearrangement. Cells were able to internalize albumin and self-assembled into chimeric 3D structures in combination with dissociated embryonic mouse kidney cells. Overall, these findings demonstrate the establishment of a robust protocol that, mimicking developmental stages, makes it possible to derive functional podocytes in vitro.http://www.sciencedirect.com/science/article/pii/S1873506116300642Induced pluripotent stem cellsDifferentiationNephron progenitorsPodocytes |
| spellingShingle | Osele Ciampi Roberto Iacone Lorena Longaretti Valentina Benedetti Martin Graf Maria Chiara Magnone Christoph Patsch Christodoulos Xinaris Giuseppe Remuzzi Ariela Benigni Susanna Tomasoni Generation of functional podocytes from human induced pluripotent stem cells Stem Cell Research Induced pluripotent stem cells Differentiation Nephron progenitors Podocytes |
| title | Generation of functional podocytes from human induced pluripotent stem cells |
| title_full | Generation of functional podocytes from human induced pluripotent stem cells |
| title_fullStr | Generation of functional podocytes from human induced pluripotent stem cells |
| title_full_unstemmed | Generation of functional podocytes from human induced pluripotent stem cells |
| title_short | Generation of functional podocytes from human induced pluripotent stem cells |
| title_sort | generation of functional podocytes from human induced pluripotent stem cells |
| topic | Induced pluripotent stem cells Differentiation Nephron progenitors Podocytes |
| url | http://www.sciencedirect.com/science/article/pii/S1873506116300642 |
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