Kidney Stem Cells Research Articles

Kidney Stem Cell Reprogramming Drives Spontaneous Hypertension via Exosomes

The spontaneously hypertensive rat (SHR) is a commonly used animal model for studying primary hypertension. However, the precise mechanism of these rats developing hypertension remains to be defined. Here, we have first demonstrated that kidney stem cell reprogramming is a driver for the development of hypertension in SHR rats. In this study, we treated 5-week-old SHR rats with WKY-kidney stem cell-derived exosomes (WKY-Exo) and 5-week-old WKY rats with SHR-Exo, respectively, via i.p. injections. Notably, WKY-Exo effectively blocked the development of hypertension and arterial stiffening in SHRs. It is fascinating that WKY rats developed hypertension and arterial stiffening due to treatment of SHR-Exo. We confirmed that both WKY- and SHR-Exo were up taken into aortic endothelial and smooth muscle cells after i.p. delivery of CD63/GFP-labelled exosomes. We demonstrated that WKY-Exo can improve vascular functions and arterial compliance of SHR rats as evidenced by preventing the development of arterial stiffness and restoring renal artery endothelial function. We also demonstrated that the vascular functions were impaired in WKY rats after SHR-Exo treatment. Furthermore, our data show that expression of Elastin, Emilin1, and Myh11 was significantly downregulated in the aorta of SHR rats, which can be repaired by WKY-Exo treatment. SHR-Exo decreased expression of Elastin and Emilin1 without affecting Myh11 in the aorta of WKY rats, suggesting that Elastin and Emilin1 deficiency could contribute to arterial stiffening leading to the development of spontaneous hypertension. Proteomic analysis reveals the enrichment of protein-protein interaction network for elastic fiber assembling, blood vessel development, and regulation of angiogenesis in WKY-Exo vs SHR-Exo. In conclusion, our data first demonstrates that kidney stem cell reprograming is a driver for the development of spontaneous hypertension in SHR rats. WKY-KSCs-derived exosome contains a proteome profile promoting arterial repair, which may provide a therapeutic means to prevent and treat primary hypertension. NIH. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Preventive interventional strategies mitigate age-associated degeneration of dorsal hippocampal neural cells in naturally ageing mice

Objectives: The objective of this study was to investigate the effectiveness and potential protective effects of various intervention strategies, such as choline and docosahexaenoic acid (Ch-DHA) supplementation, Clitoria ternatea root extract (CTR) supplements, human embryonic kidney stem cell conditioned media (HEK-CM) treatment and exposure to enriched environments (EEs), in mitigating neural cell degeneration in dorsal hippocampal subregions of naturally ageing mice brain. Materials and Methods: Twelve–fifteen-month-old CF1 male mice were subdivided (n = 6/group) into normal age-matched control, Ch-DHA supplemented, EE exposed, HEK-CM administered and CTR-supplemented groups. Neuro morphological alterations in the dorsal hippocampus were assessed using cresyl violet staining. Results: Higher neural cell degeneration was observed in the CA1–CA3 dorsal hippocampal subregions in normal ageing mice brains. Notably, interventions such as HEK-CM administration, Ch-DHA supplementation, exposure to an EE and CTR supplementation significantly reduced degeneration, particularly in the CA3 and CA2 regions. Conclusion: Ch-DHA supplementation and HEK-CM treatment are observed to significantly reduce age-dependent degeneration of dorsal hippocampal CA3 and CA2 neurons in naturally ageing mice compared to EE exposure or CTR supplementation.

Mice with renal-specific alterations of stem cell-associated signaling develop symptoms of chronic kidney disease but surprisingly no tumors.

Previously, we found that Wnt and Notch signaling govern stem cells of clear cell kidney cancer (ccRCC) in patients. To mimic stem cell responses in the normal kidney in vitro in a marker-unbiased fashion, we have established tubular organoids (tubuloids) from total single adult mouse kidney epithelial cells in Matrigel and serum-free conditions. Deep proteomic and phosphoproteomic analyses revealed that tubuloids resembled renewal of adult kidney tubular epithelia, since tubuloid cells displayed activity of Wnt and Notch signaling, long-term proliferation and expression of markers of proximal and distal nephron lineages. In our wish to model stem cell-derived human ccRCC, we have generated two types of genetic double kidney mutants in mice: Wnt-β-catenin-GOF together with Notch-GOF and Wnt-β-catenin-GOF together with a most common alteration in ccRCC, Vhl-LOF. An inducible Pax8-rtTA-LC1-Cre was used to drive recombination specifically in adult kidney epithelial cells. We confirmed mutagenesis of β-catenin, Notch and Vhl alleles on DNA, protein and mRNA target gene levels. Surprisingly, we observed symptoms of chronic kidney disease (CKD) in mutant mice, but no increased proliferation and tumorigenesis. Thus, the responses of kidney stem cells in the tubuloid and genetic systems produced different phenotypes, i.e. enhanced renewal versus CKD.

Renal regenerative capacity related to stem cell reserve in nephrectomized rats.

On the new era of stem cell therapy, the present experimental study was conducted to investigate renal regenerative capacity related to kidney stem cell reserve in different nephrectomy (Nx) models. Three- and eight-week-old rats (n = 168) were randomly divided into four groups to include control and three Nx subgroups (1/6 Nx, 1/2 Nx, and 5/6 Nx) (Fig.1). On post-Nx days 15, 30 and 60, kidney specimens were obtained to determine renal regenerative capacity. The specimens were examined with immunofluorescence. CD90/CD105 and Ki-67 expressions were determined as stem cell and cellular proliferation markers, respectively. Fig.1 Intraoperative photographs showing three different types of nephrectomies (unilateral total Nx has not been shown in 5/6 Nx group) RESULTS: CD90 and CD105 expressions were stronger in glomeruli, but Ki-67 expressions were present only in tubuli. When all Nx types and post-Nx days were considered, both 3- and 8-week-old rats undergone 5/6 Nx had the highest glomerular CD90 and CD105 double expressions. While the expressions gradually increased toward the day 60 in 3-weeks old rats, 8-week-old rats had almost stable double expressions. The strongest tubular Ki-67 expressions were seen in 5/6 Nx groups of both in 3- and 8-week-old rats. The expressions were strongest on day 15 and then gradually decreased. Ipsilateral 1/6 Nx groups had stronger Ki-67 expression than contralateral ones in both age groups. Kidneys may pose a regenerative response to tissue/volume loss through its own CD90- and CD105-related stem cell reserve which mainly takes place in glomeruli and seems to have some interactions with Ki-67-related tubular proliferative process. This response supports that kidney stem cells may have a potential to overcome tissue/volume loss-related damage.

Age-Related Changes in Kidney and Loss of Resistance to Damage: The Role of the Decrease in the Number of Kidney Progenitor Cells during Aging

Many organs undergo negative changes during aging that affect their functions and ability to regenerate. In particular, the kidneys become more susceptible to acute injury and are more likely to develop chronic kidney disease with age. One of the reasons for this may be a decrease in the number of kidney resident progenitor cells. This review addresses age-related changes that occur in the kidneys at the histological and molecular levels, including those related to the cell cycle, mitochondrial function, oxidative stress, and chronic inflammation. This review describes the available studies on resident kidney stem cells, their niches, morphology, possible markers, and the dynamics of their numbers during the aging process. The reasons for the age-related decline in renal regenerative potential are considered based on molecular and cellular mechanisms.

Exploring potential strategies to enhance memory and cognition in aging mice

Background: Aging population is rapidly expanding worldwide, and age-related cognitive impairments prove detrimental for achieving a better productive and quality of life. Lack of effective therapies for age-related cognitive impairment focuses attention on developing preventive strategies, such as nutritional interventions, cell therapies and environmental manipulations. The objective of the present study was to explore the comparative benefits of potential memory-enhancing strategies like supplementation of choline (Ch) and docosahexaenoic acid (DHA) or administration of human embryonic kidney stem cell conditioned media (HEK-CM) or exposure to environmental enrichment (EE), that attenuates cognitive impairments in aging mice. Methods: 12-month-old CF1 male mice were subdivided [n=6/group] into normal aging control (NAC), saline vehicle control (SVC), Ch-DHA, EE, heat-inactivated HEK-CM (HIHEK-CM) and HEK-CM mice. Spatial working and reference memory were assessed using an eight-arm radial maze test and cognition using a novel object recognition test (NORT). Results: Spatial memory and cognition were decreased in normal aging mice. Aged mice exposed to dietary Ch-DHA or HEK-CM showed significant enhancement in spatial learning tasks, memory and cognition compared to the same in age-matched NAC mice. Ch-DHA and HEK-CM treated mice committed significantly lesser reference memory errors and attained a higher percentage of correct choices in spatial learning and memory tasks. Moreover, on testing for cognition in NORT, significantly higher number of visits to the novel object was observed in Ch-DHA supplemented and HEK-CM administered aging mice whereas HEK-CM and EE mice groups showed significantly greater number of visits to familiar object, when compared to same in age-matched NAC and HIHEK-CM groups, respectively. Conclusion: Supplementation of Ch-DHA and HEK-CM treatment strategies have a higher potential [~ 20—30%] for enhancing spatial learning, memory and cognition in normal aged mice, whereas exposure to EE seems to enhance only their short-term memory.

Mitochondrial Toxic Effects of Antiepileptic Drug Valproic Acid on Mouse Kidney Stem Cells.

Valproic acid (VPA) is a histone deacetylase inhibitor that is used mainly as an antiepileptic and anticonvulsant drug. The side effects of VPA usually appears as hepatic injury and various metabolic disorders. On the other hand, it is rarely reported to cause kidney injury. Despite the many studies on the influence of VPA exposure on the kidneys, the specific mechanism remains unclear. This study examined the changes after VPA treatment to the mouse kidney stem cells (mKSCs). VPA triggers an increase in mitochondrial ROS, but there was no change in either mitochondrial membrane potential or the mitochondrial DNA copy number in mKSCs. The VPA treatment increased the mitochondrial complex III but decreased complex V significantly compared to the DMSO treatment as a control. The inflammatory marker (IL-6) and the expression of the apoptosis markers (Caspase 3) and were increased by VPA. In particular, the expression of the podocyte injury markers (CD2AP) was increased significantly. In conclusion, VPA exposure has adverse effects on mouse kidney stem cells.

Methods for assessment of the mitochondrial toxicity by toxicant with murine kidney stem cell

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Exploring potential strategies to enhance memory and cognition in aging mice

Background: Aging population is rapidly expanding worldwide, and age-related cognitive impairments prove detrimental for achieving a better productive and quality of life. Lack of effective therapies for age-related cognitive impairment focuses attention on developing preventive strategies, such as nutritional interventions, cell therapies and environmental manipulations. The objective of the present study was to explore the comparative benefits of potential memory-enhancing strategies like supplementation of choline (Ch) and docosahexaenoic acid (DHA) or administration of human embryonic kidney stem cell conditioned media (HEK-CM) or exposure to environmental enrichment (EE), that attenuates cognitive impairments in aging mice. Methods: 12-month-old CF1 male mice were subdivided [n=6/group] into normal aging control (NAC), saline vehicle control (SVC), Ch-DHA, EE, heat-inactivated HEK-CM (HIHEK-CM) and HEK-CM mice. Spatial working and reference memory were assessed using an eight-arm radial maze test and cognition using a novel object recognition test (NORT). Results: Spatial memory and cognition were decreased in normal aging mice. Aged mice exposed to dietary Ch-DHA or HEK-CM showed significant enhancement in spatial learning tasks, memory and cognition compared to the same in age-matched NAC mice. Ch-DHA and HEK-CM treated mice committed significantly lesser reference memory errors and attained a higher percentage of correct choices in spatial learning and memory tasks. Moreover, on testing for cognition in NORT, significantly higher number of visits to the novel object was observed in Ch-DHA supplemented and HEK-CM administered aging mice whereas HEK-CM and EE mice groups showed significantly greater number of visits to familiar object, when compared to same in age-matched NAC and HIHEK-CM groups, respectively. Conclusion: Supplementation of Ch-DHA and HEK-CM treatment strategies have a higher potential [~ 20—30%] for enhancing spatial learning, memory and cognition in normal aged mice, whereas exposure to EE seems to enhance only their short-term memory.

Gene expression studies of WT1 mutant Wilms tumor cell lines in the frame work of published kidney development data reveals their early kidney stem cell origin.

In order to get a better insight into the timing of WT1 mutant Wilms tumor development, we compared the gene expression profiles of nine established WT1 mutant Wilms tumor cell lines with published data from different kidney cell types during development. Publications describing genes expressed in nephrogenic precursor cells, ureteric bud cells, more mature nephrogenic epithelial cells and interstitial cell types were used. These studies uncovered that the WT1 mutant Wilms tumor cells lines express genes from the earliest nephrogenic progenitor cells, as well as from more differentiated nephron cells with the highest expression from the stromal/interstitial compartment. The expression of genes from all cell compartments points to an early developmental origin of the tumor in a common stem cell. Although variability of the expression of specific genes was evident between the cell lines the overall expression pattern was very similar. This is likely dependent on their different genetic backgrounds with distinct WT1 mutations and the absence/presence of mutant CTNNB1.

Inadvertent stem cell transfer demonstrating a way for tolerance induction

In this issue, Sobrino etal. present a case of hematopoietic chimerism and subsequent tolerance after isolated kidney transplantation in a pediatric patient with syndromic combined immune deficiency. This report not only highlights the chimerism approach for tolerance induction in transplantation, but it is also the first evidence of hematopoietic stem cells in human kidneys. This commentary discusses the potency and risks of the chimerism approach for tolerance induction after solid organ transplantation, especially in (pediatric) patients with immune deficiencies.

De novo SIX2 activation in human kidneys treated with neonatal kidney stem/progenitor cells

During development, nephron structures are derived from a SIX2+ stem cell population. After 36 weeks of gestation, these cells are exhausted, and no new nephrons are formed. We have previously described a non-invasive strategy to isolate and expand the native SIX2+ kidney stem cells from the urine of preterm neonates, named neonatal kidney stem/progenitor cells (nKSPC). Here, we investigated the safety and feasibility of administering nKSPC into human kidneys discarded for transplantation during normothermic machine perfusion (NMP) and evaluated the regenerative and immunomodulatory potential of nKSPC treatment. We found that nKSPC administration during NMP is safe and feasible. Interestingly, nKSPC induced the de novo expression of SIX2 in proximal tubular cells of the donor kidneys and upregulated regenerative markers such as SOX9 and VEGF. This is the first time that SIX2 re-expression is observed in adult human kidneys. Moreover, nKSPC administration significantly lowered levels of kidney injury biomarkers and reduced inflammatory cytokine levels via the tryptophan-IDO-kynurenine pathway. In conclusion, nKSPC is a novel cell type to be applied in kidney-targeted cell therapy, with the potential to induce an endogenous regenerative process and immunomodulation.

Responses of hematopoietic cells after ionizing-irradiation in anemic adult medaka (Oryzias latipes)

Purpose Hematopoietic tissues of vertebrates are highly radiation sensitive and the effects of ionizing radiation on the hematopoiesis have been studied in mammals and teleosts for decades. In this study, radiation responses in the kidney, the main hematopoietic organ in teleosts, were investigated in Japanese medaka (Oryzias latipes), which has been a model animal and a large body of knowledge has been accumulated in radiation biology. Methods Kidney, the main hematopoietic tissue of adult medaka fish, was locally irradiated using proton and carbon ion beams irradiation system of Takasaki Ion Accelerator for Advanced Radiation Application (TIARA), QST, and the effects on peripheral blood cells and histology of the kidney were investigated. Results When only kidneys were locally irradiated with proton or carbon ion beam (15 Gy), the hematopoietic cells in the irradiated kidney and cell density in the peripheral blood decreased 7 days after the irradiation in the same manner as after the whole-body irradiation with γ-rays (15 Gy). These results demonstrate that direct irradiation of the hematopoietic cells in the kidney induced cell death and/or cell cycle arrest and stopped the supply of erythroid cells. Then, the cell density in the peripheral blood recovered to the control level within 4 days and 7 days after the γ-ray and proton beam irradiation (15 Gy), respectively, while the cell density in the peripheral blood did not recover after the carbon ion beam irradiation (15 Gy). The hematopoietic cells in the irradiated kidneys temporarily decreased and recovered to the control level within 21 days after the γ-ray or proton beam irradiation (15 Gy), while it did not recover after the carbon ion beam irradiation (15 Gy). In contrast, the recovery of the cell density in the peripheral blood delayed when anemic medaka were irradiated 1 day after the administration of phenylhydrazine. With and without γ-ray irradiation, a large number of hematopoietic cells was still proliferating in the kidney 7 days after the anemia induction. Conclusions The results obtained strongly suggest that the hematopoietic stem cells in medaka kidney prioritize to proliferate and increase peripheral blood cells to eliminate anemia, even when they are damaged by high-dose irradiation.

Methods to measure the mitochondrial toxicity in mouse kidney stem cell by toxicant

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Potential Strategies for Kidney Regeneration With Stem Cells: An Overview.

Kidney diseases are a major health problem worldwide. Despite advances in drug therapies, they are only capable of slowing the progression of kidney diseases. Accordingly, potential kidney regeneration strategies with stem cells have begun to be explored. There are two different directions for regenerative strategies, de novo whole kidney fabrication with stem cells, and stem cell therapy. De novo whole kidney strategies include: 1) decellularized scaffold technology, 2) 3D bioprinting based on engineering technology, 3) kidney organoid fabrication, 4) blastocyst complementation with chimeric technology, and 5) the organogenic niche method. Meanwhile, stem cell therapy strategies include 1) injection of stem cells, including mesenchymal stem cells, nephron progenitor cells, adult kidney stem cells and multi-lineage differentiating stress enduring cells, and 2) injection of protective factors secreted from these stem cells, including growth factors, chemokines, and extracellular vesicles containing microRNAs, mRNAs and proteins. Over the past few decades, there have been remarkable step-by-step developments in these strategies. Here, we review the current advances in the potential strategies for kidney regeneration using stem cells, along with their challenges for possible clinical use in the future.

Adult Mouse Kidney Stem Cells Orchestrate the De Novo Assembly of a Nephron via Sirt2‐Modulated Canonical Wnt/β‐Catenin Signaling (Adv. Sci. 15/2022)

De Novo Generation of Kidney Organoids Using Adult Kidney Stem Cells Generation of kidney organoids using autologous kidney stem cells represents an attractive strategy for treating and potentially replacing the failing kidneys. In article number 2104034, Xiaobin Han and Zhongjie Sun report a previously unidentified adult kidney Sca1+ Oct4+ stem cells. Interestingly, culturing these stem cells led to de novo generation of self-organizing nephrons and kidney-like structures. This finding provides the first evidence that kidney stem cells are capable of de novo generating kidney organoids. This finding also offers a new opportunity for studying mammalian kidney development and personalized kidney regeneration.

Adult mouse kidney stem cell‐derived exosomes navigate nephron repair and regeneration in aged mice

The physiological function of kidney stem cell‐derived extracellular vesicles remains elusive. Most recently, we isolated kidney stem cells (KSCs) from adult mouse kidneys and demonstrated that KSCs‐derived exosomes normalized blood pressure and improved kidney functions in hypertensive aged mice. To study if KSCs‐derived exosomes play a role in nephron repair and regeneration, we treated aged mice with CD63‐GFP labelled KSC‐exosome (CD63/GFP KSC‐Exo) (~1.0×1011particles/mouse) and traced CD63/GFP KSC‐Exo in the kidney. We found that KSC‐exosomes were mainly detected in the proximal and distal tubular cells near the damaged glomerulus in cortex area of the aging kidney. BrdU tracing study showed that BrdU positive cells were mainly detected in the glomerulus and co‐located with Ki67 cell proliferation marker in the kidney of KSC‐exosome treated mice. KSC‐exosome treatment also increased numbers of Sca1+cells surrounding the glomerulus area of aging kidney. Interestingly, the glomerular filtration rate (GFR) was significantly increased in KSC‐exosome‐treated aged mice, suggesting improved glomerular function. To explore the mechanisms how KSC‐exosome improves kidney functions, we performed proteome analysis of KSC‐exosomes by mass spectrometry. We found several pathways that involve in the kidney development were upregulated in the KSC‐exosome from adult mice compared to that of aged mice. Notably, we further identified that FBLN2, an extracellular matrix protein, plays an important role in aging‐associated hypertension. We showed that expression of FBLN2 was downregulated in the kidney of aged mice and recombinant FBLN2 (rFBLN2) treatment normalized blood pressure and improved GFR of hypertensive aged mice. Furthermore, we found that FBLN2 is mainly located in the basement membrane of glomerulus (GBM). The thickness of GBM was increased in aged mice determined by TEM. KSC‐exosome or rFBLN2 treatment reduced the thickness of GBM, suggesting that FBLN2 may play an important role in maintaining the integrity of GBM and kidney functions. In conclusion, our finding provided the first evidence that the adult mouse kidney stem cell‐derived exosomes play an important role in nephron repair and regeneration partly via FBLN2. KSC‐exosome treatment may be a novel therapeutic strategy for hypertension‐associated kidney injuries in aging.

Adult mouse kidney stem cells orchestrate the de novo assembly of a nephron via Sirt2‐ modulated canonical Wnt/β‐catenin signaling

End‐stage renal disease affects an estimated 2.4 million people annually worldwide. With cases increasing 8% every year, regenerative strategies are needed to help alleviate, and potentially replace, the increasing demand for kidney transplants. An important question remains whether stem cells reside in the adult mammalian kidney. Here we developed a methodology for isolation of kidney stem cells from adult mouse kidney by using Sca1 and Oct4 stem cell markers. Using these cells, we generated three types of kidney‐like structures. First, we observed the assembly of self‐organizing three‐dimensional kidney organoids. These kidney organoids show characteristic of kidney structures and contain podocytes and endothelial cells that form networks of capillary loop‐like structures. We also observed the de novogeneration of a self‐organizing nephron (and its associated collecting duct) from monolayer cultures. Comma‐ and S‐shaped bodies that expressed activated Notch1 were present during this formation, suggesting that Notch signaling played a key role in driving nephrogenesis. This is consistent with our current knowledge of nephron formation in the mouse embryo. Furthermore, we observed the differentiation of monolayer cells into functionally mature tubules and self‐organizing kidney‐shaped structures that selectively endocytose dextran. Finally, we demonstrated that expression of Sirt2 was upregulated, which modulated the dynamics of canonical Wnt/β‐catenin signaling during kidney organoid development. Altogether, our observations provide the first evidence for the existence of stem cells in the adult mouse kidney. These findings offer powerful tools for future applications, including drug screening, disease modelling, cell therapy, and kidney regeneration.

Isolation of adult rat kidney derived stem cells and differentiation into podocyte-like cells

Studies have shown that adult stem cells can be isolated from different organs. However, no agreement has yet been reached on the identification of adult kidney stem cells. In this study, we have shown that the adult rat kidney contains a population of stem cells that are isolatable and, under suitable culture conditions, these stem cells can differentiate into podocyte. Six adult Wistar rats were used in the present study. Their kidneys were chopped and exposed to collagenase I. Then, with passing through 100 μm mesh, single cells in filtered fraction were cultured in a proliferation medium. After 4 passages, the cells were analyzed by flow cytometry, differentiation and gene expression. Morphological analyses revealed that isolated cells are spindle-shaped, and can differentiate into osteocytes and adipocytes. Flow cytometry demonstrated that these cells expressed cell surface markers CD44, CD90, CD133, c-kit, Pax- 2, Oct4, sca-1 and vimentin significantly. Also, the high expression of Wt-1 and Wnt-4 genes was seen in these cells. Moreover, expression of synaptopodin and podocalyxin genes showed differentiation of stem cells into podocyte cells. Our findings demonstrate that isolated cells from the kidney of adult rats are mesenchymal stem cells or progenitor cells and can differentiate to podocyte-like cells.

Adult Mouse Kidney Stem Cells Orchestrate the De Novo Assembly of a Nephron via Sirt2-Modulated Canonical Wnt/β-Catenin Signaling.

Generation of kidney organoids using autologous kidney stem cells represents an attractive strategy for treating and potentially replacing the failing kidneys. However, whether adult mammalian kidney stem cells have regenerative capacity remains unknown. Here, previously unidentified adult kidney Sca1+ Oct4+ stem/progenitor cells are isolated. Interestingly, culturing these cells leads to generation of kidney‐like structures. First, the assembly of self‐organizing 3D kidney‐like structures is observed. These kidney organoids contain podocytes, proximal tubules, and endothelial cells that form networks of capillary loop‐like structures. Second, the differentiation of kidney stem cells into functionally mature tubules and self‐organizing kidney‐shaped structures in monolayer culture that selectively endocytoses dextran, is shown. Finally, the de novo generation of an entire self‐organizing nephron from monolayer cultures is observed. Mechanistically, it is demonstrated that Sirt2‐mediated canonical Wnt/β‐catenin signaling is critical for the development of kidney organoids. Thus, the first evidence is provided that the adult mouse kidney stem cells are capable of de novo generating kidney organoids.