使用 Seahorse 分析类器官生物能量学的方案。

Molecular Cancer Research, Center for Molecular Medicine, University Medical Center Utrecht, Heidelberglaan 100, 3584 CG Utrecht, the Netherlands.

Oncode Institute.

STAR Protoc. 2021 Mar 18;2(1):100386. doi: 10.1016/j.xpro.2021.100386. eCollection 2021 Mar 19.

Addressing bioenergetics is key to evaluate the impact of metabolism on the regulation of biological processes and its alteration in disease. Organoids are grown self-organizing structures derived from healthy and diseased tissue that recapitulate with high fidelity the tissue of origin. Bioenergetics is commonly analyzed by Seahorse XF analysis. However, its application to organoid studies is technically challenging. Here, we share our in-house optimized protocols to examine organoid bioenergetics in response to drugs, gene knockdown, or to characterize the metabolism of specific cell types. For complete details on the use and execution of this protocol, please refer to Ludikhuize et al. (2020).

解决生物能量学问题是评估代谢对生物过程调控的影响及其在疾病中的变化的关键。类器官是从健康和患病组织中生长出来的自组织结构，能够高度忠实地再现组织的起源。生物能量学通常通过 Seahorse XF 分析进行分析。然而，将其应用于类器官研究在技术上具有挑战性。在这里，我们分享我们内部优化的方案，以研究类器官生物能量学对药物、基因敲低的反应，或用于表征特定细胞类型的代谢。有关该方案使用和执行的完整详细信息，请参阅 Ludikhuize 等人（2020 年）。