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Abstract: TH-PO635

Development and Characterization of Kidney Micro-Organoids from PSCs to Facilitate Kidney Cell Scale-Up

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 501 Development, Stem Cells, and Regenerative Medicine: Basic


  • Kumar, Santhosh V, Murdoch Children's Research Institute, Parkville, New South Wales, Australia
  • Er, Pei Xuan, Murdoch Children's Research Institute, Parkville, New South Wales, Australia
  • Lawlor, Kynan T., Murdoch Children's Research Institute, Parkville, New South Wales, Australia
  • Combes, Alexander N., Murdoch Children's Research Institute, Parkville, New South Wales, Australia
  • Little, Melissa H., Murdoch Children's Research Institute, Parkville, New South Wales, Australia

Group or Team Name

  • Kidney development, disease and regeneration

The directed differentiation of human pluripotent stem cells (hPSCs) has enabled the generation of kidney organoids with potential applications in disease modelling, drug screening and regenerative medicine. The approach is expensive and faces limitations to long-term culture. As a result, novel cost-effective techniques are needed to enable scale-up of kidney cells types in vitro and facilitate higher throughput screening approaches.


To generate kidney micro-organoids, hPSCs were differentiated to intermediate mesoderm using 7uM CHIR for 4 days and FGF9 until 7 days in 2D monolayer culture. On day 7 cells were dissociated using EDTA and allowed to form cell aggregates via low-speed swirling using an E6 media containing a cocktail of growth factors. Within 24hr, cell aggregates were transformed into cell aggregates which, after a further 12 to 18 days in suspension culture, formed complex kidney micro-organoids within a single culture flask.


Mature kidney micro-organoids revealed the presence of intact nephron segments, including podocytes (NPHS1+), proximal (LTL+) and distal (ECAD+) nephron epithelium and collecting duct (ECAD+GATA3+) segments with distinct lumens. Each micro-organoid contains only 6-10 nephrons. As a result, imaging through an entire organoid is feasible. 10x Chromium single-cell RNA sequencing analysis identified the presence of 11 distinct kidney cell types, both confirming the presence of anticipated renal cell types and revealing a reduced level of surrounding stroma compared to our previously described protocol (Takasato et al, 2015). With this novel method, a starting population of 4.5 million cells (D7+0) was expanded to 150-200 million kidney cells (40 fold) within 20 days, providing a platform for the economical production of kidney cells for various biological applications. Treatment of organoids with Adriamycin induced podocyte cell death and decreased podocyte specific gene expression.


This is a cost-effective method for the generation of large numbers of hPSC-derived kidney cells. As a result, this technique will improve the feasibility of drug toxicity screening and regenerative cell therapy.