Protocol to analyze bioenergetics in single human induced-pluripotent-stem-cell-derived kidney organoids using Seahorse XF96

STAR Protocols, vol. 4, pp. 101999

Abstract

Metabolic derangement is a key culprit in kidney pathophysiology. Organoids have emerged as a promising in vitro tool for kidney research. Here, we present a fine-tuned protocol to analyze bioenergetics in single human induced-pluripotent-stem-cell (iPSC)-derived kidney organoids using Seahorse XF96. We describe the generation of self-organized three-dimensional kidney organoids, followed by preparation of organoids for Seahorse XF96 analysis. We then detail how to carry out stress tests to determine mitochondrial and glycolytic rates in single kidney organoids.

Authors 8

1. Verónica Miguel corresponding Aachen Institute of Experimental Medicine and Systems Biology

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Experimental Medicine and Systems Biology, RWTH Aachen University Hospital, Aachen, Germany. Electronic address: vmiguh00@gmail.com

   Institute of Experimental Medicine and Systems Biology, RWTH Aachen University Hospital, Aachen, Germany

2. Katharina C. Reimer Aachen Institute of Experimental Medicine and Systems Biology Institute for Biomedical Technologies Department of Cell Biology Department of Nephrology and Hypertensive Disorders Clinical Immunology (Medical Clinic II) Institute for Biomedical Technologies Department of Cell Biology

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Experimental Medicine and Systems Biology, RWTH Aachen University Hospital, Aachen, Germany; Institute for Biomedical Technologies, Department of Cell Biology, RWTH Aachen University, Aachen, Germany; Department of Nephrology and Hypertensive Disorders, Rheumatology, and Clinical Immunology (Medical Clinic II), University Hospital RWTH Aachen, Aachen, Germany

   Department of Nephrology and Hypertensive Disorders, Rheumatology, and Clinical Immunology (Medical Clinic II), University Hospital RWTH Aachen, Aachen, Germany

   Institute for Biomedical Technologies, Department of Cell Biology, RWTH Aachen University, Aachen, Germany

   Institute of Experimental Medicine and Systems Biology, RWTH Aachen University Hospital, Aachen, Germany

3. Anna Katharina Galyga Aachen Institute for Biomedical Technologies Department of Cell Biology

   RWTH Aachen University

   Affiliation as printed

   Institute for Biomedical Technologies, Department of Cell Biology, RWTH Aachen University, Aachen, Germany

4. Jitske Jansen Aachen Institute of Experimental Medicine and Systems Biology

   Radboud University Nijmegen · Radboud University Medical Center · Radboud Institute for Molecular Life Sciences · Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Experimental Medicine and Systems Biology, RWTH Aachen University Hospital, Aachen, Germany; Department of Pathology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands

   Department of Pathology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands

   Institute of Experimental Medicine and Systems Biology, RWTH Aachen University Hospital, Aachen, Germany

5. Julia Möllmann Aachen Department of Internal Medicine I (Cardiology)

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Department of Internal Medicine I, Cardiology, University Hospital RWTH Aachen, Aachen, Germany

6. Luisa H. Meyer Aachen Department of Internal Medicine I (Cardiology)

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Department of Internal Medicine I, Cardiology, University Hospital RWTH Aachen, Aachen, Germany

7. Rebekka K. Schneider Aachen Institute for Biomedical Technologies Department of Cell Biology

   RWTH Aachen University

   Affiliation as printed

   Institute for Biomedical Technologies, Department of Cell Biology, RWTH Aachen University, Aachen, Germany

8. Rafael Kramann corresponding Aachen Institute of Experimental Medicine and Systems Biology Department of Nephrology and Hypertensive Disorders Clinical Immunology (Medical Clinic II)

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Experimental Medicine and Systems Biology, RWTH Aachen University Hospital, Aachen, Germany; Department of Nephrology and Hypertensive Disorders, Rheumatology, and Clinical Immunology (Medical Clinic II), University Hospital RWTH Aachen, Aachen, Germany. Electronic address: rkramann@gmx.net

   Department of Nephrology and Hypertensive Disorders, Rheumatology, and Clinical Immunology (Medical Clinic II), University Hospital RWTH Aachen, Aachen, Germany

   Institute of Experimental Medicine and Systems Biology, RWTH Aachen University Hospital, Aachen, Germany