Subcellular lipidomics is a novel field of research that requires the careful combination of several pre-analytical and analytical steps. To define a reliable strategy for mitochondrial lipid profiling, we performed a systematic comparison of different mitochondria isolation procedures by western blot analyses and comprehensive high-resolution lipidomics. Using liver-derived HepG2 cells, we com...

Mitochondria regulate critical components of cellular function via ATP production, reactive oxygen species production, Ca(2+) handling and apoptotic signaling. Two classical methods exist to study mitochondrial function of skeletal muscles: isolated mitochondria and permeabilized myofibers. Whereas mitochondrial isolation removes a portion of the mitochondria from their cellular environment, my...

Mitochondria are complex organelles constantly undergoing processes of fusion and fission, processes that not only modulate their morphology, but also their function. Yet the assessment of mitochondrial function in skeletal muscle often involves mechanical isolation of the mitochondria, a process which disrupts their normally heterogeneous branching structure and yields relatively homogeneous s...

AIM The aim of this study was to evaluate and compare three available methods for mitochondrial isolation from a human cell line to predict the best method for each probable application. BACKGROUND Organelle isolation is gaining importance in experimental laboratory settings. Mitochondrial dysfunction may affect tumorgenesis process. There are some evidences that transplantation of healthy, i...

1. An improved type of ground glass homogenizer for soft tissues has been described which brings about a high degree of cell disruption and liberation of nuclei without causing appreciable damage to mitochondria. The gentleness and effectiveness of the new homogenizer in respect to isolation of mitochondria have been ascertained by comparing the ATP-ase activities of mitochondria isolated in 0....

Mitochondria play key roles in energy metabolism and cell physiology. The properties and metabolic capacity of mitochondria in suspension cultures of mammalian cells have been seldom studied despite their importance for the production of recombinant proteins. Current metabolic studies of mammalian cells generally examine the cells as a whole, regardless of the compartmentalization of metabolism...

A number of parameters associated with the isolation of plant mitochondria were carefully examined in the hope of optimizing the isolation technique. Dark-grown mung bean (Phaseolus aureus var. Jumbo) hypocotyls were used as the source of mitochondria. Necessary conditions for the isolation of tightly coupled plant mitochondria include: (a) gentle tissue disruption; (b) rigorous exclusion of co...

Two methods of preparing a mitochondrial fraction from beef brain cortex are described. Data are presented on the rate of oxidation of substrates, P/O and respiratory control ratios, cholinesterase activity, and DNA content. Electron micrographs of isolated mitochondria and mitochondria in situ are shown. Comparisons are drawn between these preparations and mitochondria prepared in 0.25 M sucro...