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Kidney Week

Abstract: SA-PO551

Growth, Maturation, and Light Sheet Microscopy of Autologous Induced Pluripotent Stem Cell-Derived Kidney Organoids in Tunable Gelatin Methacrylate (GelMA) Hydrogels

Session Information

  • Bioengineering
    October 26, 2024 | Location: Exhibit Hall, Convention Center
    Abstract Time: 10:00 AM - 12:00 PM

Category: Bioengineering

  • 400 Bioengineering

Authors

  • Krupa, Ivan, University College Dublin, Dublin, Ireland
  • Clerkin, Shane, University College Dublin, Dublin, Ireland
  • Davis, Jessie, University College Dublin, Dublin, Ireland
  • Knezevic, Sara, University College Dublin, Dublin, Ireland
  • Lees, Robert, Science and Technology Facilities Council, Swindon, Swindon, United Kingdom
  • Brougham, Dermot F., University College Dublin, Dublin, Ireland
  • Crean, John, University College Dublin, Dublin, Ireland
Background

Recent advances in the generation of hiPSC derived kidney organoids have successfully established their use as a novel disease-modelling platform. Here we describe the utility of gelatin methacryloyl (GelMA) hydrogels as support matrices for kidney organoids in which manipulation of biophysical parameters like stiffness and shear flow acts as a mechanism for control of cell composition and structure.

Methods

Kidney organoids were generated from hiPSCs using established methods (Treacy et al., Bioactive Matertials, 2023). Light sheet fluorescence microscopy (LSFM) imaging was conducted on a Leica SP8 Digital Light sheet microscope. Hydrogel rheology was conducted using the Anton Parr Physica MCR301 rheometer. For experiments investigating the effects of shear flow via the use of an orbital shaker system, a modified version of a protocol published by Przepiorski et al., (2021) was utilised in conjunction with GelMA hydrogel encapsulation of kidney organoids.

Results

Rheological measurements show that chemically crosslinked GelMA hydrogels retain their mechanical storage moduli over a prolonged period of time in culture. Therefore, this biomaterial provides a means of studying the effects of a constant storage modulus of the surrounding matrix on the growth of organoids.We have recently evaluated the utility of LSFM as a technique for rapid 3D imaging of organoid samples and a means for characterising the effects of the biophysical microenvironment on kidney organoid generation. In this pilot study, we imaged the development of MAFB+ve cells, representing developing podocytes, within kidney organoids grown in GelMA, with live low-phototoxicity light sheet imaging. We also observed CD31+ve endothelial cells in GelMA supported kidney organoids with fixed and cleared light sheet imaging. The extent of the vascularisation observed was comparable to that previously described (Ryan et al., Developmental biology, 2021).

Conclusion

Overall, our study suggests that modification of biophysical properties such as matrix stiffness and shear flow can be used as an instructive cue to modify kidney organoid maturation. With this work we hope to inform the development of increasingly physiologically authentic kidney organoids for the advancement of 3D models of disease and drug responses.

Funding

  • Government Support – Non-U.S.