3.3. Using pluripotent stem cell-derived kidney organoids to model kidney injury

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1,2Theresa Chow, 2Jennifer Whiteley, 1,2,3Ian Rogers

1 Department of Physiology, Faculty of Medicine, University of Toronto, Ontario, Canada; 2 Lunenfeld Tanenbaum Research Institute, Mount Sinai Hospital, Ontario, Canada; 3 Department of Obstetrics and Gynecology, Faculty of Medicine, University of Toronto, Ontario, Canada

The conventional approach for drug and toxicity screening involves culturing primary cells or cell lines on 2D surfaces and treating confluent cells with drugs. This approach is limited by primary tissue availability, cell de-differentiation, and/or cell senescence. Embryonic stem cells are pluripotent stem cells capable of self-renewal and differentiation to somatic cells of all three germ layers. These properties make pluripotent stem cells great candidates to use for drug and toxicity screening. Here, we show that mouse embryonic stem cells can be differentiated to renal progenitors, and these renal progenitors can be coaxed to form kidney organoids. Our differentiation protocol differentiates embryonic stem cells to mesoderm, intermediate mesoderm, and progenitors in a step wise fashion. Similar to the embryonic kidney, our progenitors express markers of metanephric mesenchyme (Six2, Wt1, Pax2), ureteric bud (cRet, Gata3, Hoxb7), stromal progenitors (Foxd1), and endothelial progenitors (CD34). When cultured as a cell aggregate on an air-liquid interface, our progenitors give rise to kidney organoids containing mature tubules. The organoid and the tubular structures within it are supported by extracellular matrix proteins (collagen IV, laminin, fibronectin) produced by the organoid itself. The kidney organoids respond to cisplatin, a nephrotoxic drug, in a dose-dependent manner by upregulating kidney injury markers Caspase 3, IL18, IL6 and IL8. The addition of 5 mM N-acetyl-cysteine (NAC) or 1 mM dimethylthiourea (DMTU) can alleviate cisplatin-induced kidney injury. Together, our data shows that pluripotent stem cell-derived kidney oragnoids can be used as a platform for drug and nephrotoxicity screens.