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

利用 Seahorse XF96 分析单个人类诱导多能干细胞衍生肾脏类器官的生物能量学方案

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作者:Verónica Miguel ,Katharina Charlotte Reimer ,Anna Katharina Galyga ,Jitske Jansen ,Julia Möllmann ,Luisa Meyer ,Rebekka K Schneider ,Rafael Kramann

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. Keywords: Cell Differentiation; Metabolism; Organoids; Stem Cells.

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