Generation of functional podocytes from human induced pluripotent stem cells

Abstract

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.

A CRISP(e)R view on kidney organoids allows generation of an induced pluripotent stem cell–derived kidney model for drug discovery

“…In an effort to generate tools for both drug discovery and drug safety within the area of kidney disease, we aimed to develop an in vitro model for glomerular health using hiPSCs. To date, some recent progress has been made toward this end, [1][2][3][4][5]28 but there are still no established functional tools mimicking the human situation in the screening format required for drug discovery.…”

“…The induced pluripotent stem cell (iPSC) technology has made it possible to access and modify cells of a wide variety of origins that previously have been difficult or nearly impossible to study. Within the field of nephrology, some recent advances have been made in creating 3-dimensional (3D) organoid cultures, [1][2][3][4][5] but there are still limited functional tools for drug discovery available.…”

Nephrotic syndrome in a dish: recent developments in modeling in vitro

“…Urinary renal progenitor cells have the potential to be used as a non-invasive source for studying personalized genetic kidney disease, but the number of progenitor cells present in urine varies and hampers the success rate of urine-derived progenitor and associated podocyte cultures. To overcome this hurdle, the use of (patient-derived) induced pluripotent stem cells (iPSC) and differentiation towards podocytes is a promising approach [21][22][23][24]. Tanigawa and co-workers successfully reprogrammed skin fibroblasts isolated from a newborn diagnosed with congenital NS into iPSC [12].…”

“…2). Using either 2D or 3D culture approaches, and varying the choice and/or timing of growth factors, result in the induction of one renal cell type (e.g., podocyte-like cells) [15,21,55,56] or in self-organizing kidney organoids consisting of segmented nephrons [22,23,[57][58][59]. These protocols are extensively reviewed by Soo et al [60].…”

Kidney Organoids: A Translational Journey

“…Currently, there are several studies that have reported the differentiation of hPSCs into cells of the kidney lineage [3–7, 34, 35, 64]. Kidneys consist of many different cell types in the nephron, vasculature and interstitial compartments.…”