We are pleased to share that in our lab, a novel method was developed for isolating mitochondria specifically from rodent cardiomyocytes, ensuring high purity and preservation of their structural and functional integrity. Unlike conventional techniques based on harsh mechanical homogenization, our approach uses gentle mechanical disruption, allowing for the recovery of mitochondria from both subsarcolemmal and interfibrillar regions of the cell. This method significantly improves mitochondrial yield while minimizing contamination from other cell types. The integrity and functionality of the isolated mitochondria were confirmed using advanced techniques, including transmission electron microscopy, high-resolution respirometry, and fluorescence-based measurements of membrane potential. The mitochondria exhibited robust respiratory activity and remained suitable for demanding applications such as single-channel patch-clamp electrophysiology and mitochondrial transplantation into cultured cells. Overall, the protocol is rapid, efficient, and versatile, providing high-quality mitochondrial preparations for a wide range of biomedical and pharmacological studies.
Joanna Lewandowska, Barbara Kalenik, Adam Szewczyk, Antoni Wrzosek (2026) Rapid protocol for mitochondria isolation from cardiomyocytes employing cell strainer-based procedure. bioRxiv ; doi: 10.64898/2026.04.02.716092
