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 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.
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy; Roche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
Osele
O
Ciampi
Roche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
Roberto
R
Iacone
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy.
Lorena
L
Longaretti
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy.
Valentina
V
Benedetti
Roche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
Martin
M
Graf
Roche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
Maria Chiara
MC
Magnone
Roche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
Christoph
C
Patsch
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy.
Christodoulos
C
Xinaris
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy; Unit of Nephrology and Dialysis, Azienda Socio Sanitaria Territoriale (ASST) Papa Giovanni XXIII, 24127 Bergamo, Italy; Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy.
Giuseppe
G
Remuzzi
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy. Electronic address: ariela.benigni@marionegri.it.
Ariela
A
Benigni
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy.
Susanna
S
Tomasoni
D000185:Actinin / Q000235:genetics / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D058732:Embryoid Bodies / Q000378:metabolism / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D059785:Karyotype; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D050199:Podocytes / Q000166:cytology* / Q000378:metabolism; D016708:Synaptophysin / Q000235:genetics / Q000378:metabolism
D015820:Cadherins; D008565:Membrane Proteins; C504260:SYNPR protein, human; D016708:Synaptophysin; C113267:nephrin; D000185:Actinin
Differentiation; Induced pluripotent stem cells; Nephron progenitors; Podocytes