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Abstract: SA-PO324

Highly Parallel Production of Designer Kidney Organoids by Mosaic Patterning of Progenitors

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 600 Development, Stem Cells, and Regenerative Medicine


  • Porter, Catherine M., University of Pennsylvania Department of Bioengineering, Philadelphia, Pennsylvania, United States
  • Hughes, Alex, University of Pennsylvania Department of Bioengineering, Philadelphia, Pennsylvania, United States

Group or Team Name

  • Hughes Lab.

Kidney organoids derived from human induced pluripotent stem cells (iPSCs) are a promising approach for congenital and adult disease modeling, regenerative medicine, and gaining fundamental understanding of organogenesis. However, organoids are limited in their production scale, reproducibility, and physiologic function and structure, posing a considerable barrier to their application. Thus, an important open question is the extent to which engineering control over culture conditions can guide organoids toward specific outcomes in composition and organization. Here, we study this by precisely controlling iPSC-derived kidney progenitor cell numbers and ratios in microwell-format organoid cultures.


We began by adapting transient 2D cell patterning technology to a microwell format suited for transition to long-term 3D organoid culture. We first applied our validated culture system to study the effect of initial nephron progenitor (NP) number on organoid morphogenesis. Since reciprocal interactions between NPs and ureteric bud (UB) tip cells influence nephron endowment during kidney organogenesis, a major predictor of kidney disease, we next examined how modulating ratios of these cells affects organoid morphogenesis.


In our NP-only organoids, we discovered that throughout culture, organoid sizes directly depend on the number of initial NPs. Moreover, larger nephron organoids display a shift in final composition, with a significant increase in proximal tubule proportion from 40% to 61% from 2D cell pattern size diameters 200 to 500 µm.

Preliminarily, we have found that regardless of cell ratio, transition of 2D mosaic cell patterns of NPs and UB tip cells to 3D suspension culture facilitates cell sorting and discrete compartment formation early in organoid development, which resembles physiologic organization. Furthermore, UB tip cells and NPs synergistically upregulate stromal cell populations.


Here, we contribute an integrated cell patterning and long-term culture platform that decouples organoid size from well geometry, enables 3D suspension culture, and maintains discrete organoid cultures. Our approach makes significant advances in organoid homogeneity relevant to drug screens and modeling of development and diseases as well as improved control over tissue interface formation crucial to engineered morphogenesis efforts.


  • NIDDK Support