Integrative analysis of mitochondrial metabolic dynamics in reprogramming human fibroblast cells

Abstract

Summary: Mitochondrial dynamics play critical roles in both tissue homeostasis and somatic cell reprogramming. Here, we provide integrated guidance for assessing mitochondrial function and dynamics while reprogramming human fibroblasts via an integrated analysis approach. This protocol includes instructions for mitochondrial metabolic analysis in real time and flow cytometry-based assessment of mitochondrial mass and membrane potential. We also describe a protocol for quantification of mitochondrial network and key metabolites.For complete details on the use and execution of this protocol, please refer to Cha et al. (2021). : Publisher's note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Cell culture
• Flow Cytometry/Mass Cytometry
• Cell-based Assays
• Metabolism
• Microscopy