Separation of mitochondria in cancer cell-derived cells and metabolic assessment

摘要

The cell membrane destruction process, which takes the longest time in the existing method, was shortened by using a bead beater. With this requirement, it is possible to destroy the cell membrane in 5 seconds of 24 samples compared to the conventional method, which requires about 5 minutes per sample. In addition, it was possible to obtain sufficient mitochondria to the extent possible for Western blot experiments by extracting metabolites, lipid bodies, and proteins using this. It was confirmed that a fraction of high purity was obtained as a result of using Western blot and scanning electron microscope, which are conventional methods of checking purity. However, as a result of the analysis, ER could not be completely removed from mitochondria, which may cause bias in mitochondrial metabolism and lipid analysis results. Nevertheless, the results presented in this study are considered to be the most efficient method to date, overcoming representative obstacles raised in existing mitochondrial separation and metabolism studies.