A simple bioreactor-based method to generate kidney organoids from pluripotent stem cells

Abstract

SummaryKidney organoids generated from human pluripotent stem cells have the potential to revolutionize how kidney development and injury are studied. Current protocols are technically complex and suffer from poor reproducibility and high reagent costs restricting scalability. To overcome these issues, we have established a simple, inexpensive and robust method to grow kidney organoids in bulk from human induced pluripotent stem cells. Our organoids develop tubular structures by day (d) 8 and show optimal tissue morphology at d14. A comparison with fetal human kidney suggests that d14 organoid renal structures most closely resemble ‘capillary loop’ stage nephrons. We show that deletion of HNF1B, a transcription factor linked to congenital kidney defects, interferes with tubulogenesis, validating our experimental system for studying renal developmental biology. Taken together, our protocol provides a fast, efficient and cost-effective method for generating large quantities of human fetal kidney tissue, enabling the study of normal and aberrant human renal development.

Modeling acute kidney injury in kidney organoids with cisplatin

“…All work was carried out with the approval of Human Participants Ethics Committees (UAHPEC 8712 and HDEC 17/NTA/204) and biosafety. iPSC maintenance and kidney organoid generation were performed as described (13). All experiments were performed using organoids generated from the MANZ-2 iPSC line (13).…”

“…We have developed a novel kidney organoid protocol that uses a simple, cost-effective method to generate large numbers of organoids, making it ideally suited for injury modeling and drug development (13). Here, we used these kidney organoids to analyze the effects and specificity of cisplatin-induced AKI.…”

Kidney organoid reproducibility across multiple human iPSC lines and diminished off target cells after transplantation revealed by single cell transcriptomics

“…Technologies to generate kidney organoids from human induced pluripotent stem cells (iPSC) (Cruz et al, 2017a;Forbes et al, 2018a;Freedman et al, 2015;Morizane and Bonventre, 2017a;Morizane et al, 2015;Przepiorski et al, 2018;Taguchi and Nishinakamura, 2017;Taguchi et al, 2014;Takasato et al, 2015aTakasato et al, , 2016aTanigawa et al, 2018a) provide a promising avenue to further help advance our understanding of disease mechanisms and expedite therapeutic development.…”

From organoids to transplantable artificial kidneys

“…Takasato et al [5,6] developed a relatively simple protocol, while the method of Morizane et al [7] required a shorter culture time. Recently, Przepiorski et al [8] reported a simple and cost-effective protocol using a bioreactor. These protocols share certain features but also have some distinct aspects.…”

“…These protocols share certain features but also have some distinct aspects. Although the similarities and differences between the nephron progenitors and nephrons induced by these different protocols need to be determined, the induced proximal renal tubules have been shown to carry out some functions, including reabsorption of low-molecular-weight dextran [6,8] and selective cell death in response to nephrotoxic drugs [6,7].…”