Centrifugation‐Free Magnetic Isolation of Functional Mitochondria Using Paramagnetic Iron Oxide Nanoparticles

Abstract

Subcellular fractionation techniques are of tremendous importance in the field cellular biology, drug development, and many others. With the emergence of organelle targeted nanoparticle (NP) platforms, it is getting increasingly important to isolate target organelles in order to determine localization and activity of these agents. Mitochondria targeted NPs attracted attention of researchers around the globe since dysfunctions of this organelle can result in a wide range of diseases. Conventional mitochondria isolation methods involve high-speed centrifugation. The inherent problem associated with high speed centrifugation-based isolation of NP-loaded mitochondria using these conventional methods can arise from settling down of NPs along with the organelle irrespective of whether these NPs are truly associated with mitochondria or not. We report development of a mitochondria-targeted paramagnetic iron oxide nanoparticle, Mito-magneto, that enables isolation of mitochondria under the influence of a magnetic field. The mitochondria isolated using this centrifugation-free method are not only intact, pure and respiration active but also eliminates artifacts that are typically associated with isolation of NP-loaded mitochondria using centrifugation.

An Improved Method for Preparation of Uniform and Functional Mitochondria from Fresh Liver

“…Besides equilibrium density gradient ultracentrifugation, other physical methods have been studied to improve the purity of mitochondrial preparations. For example, Banik and Dhar12 isolated functional mitochondria using paramagnetic iron oxide nanoparticles. Although purification was achieved, the nanoparticles could not be separated from the mitochondria after isolation, which might limit the value of this method.…”

Dual-Targeted Synthetic Nanoparticles for Cardiovascular Diseases

“…Thus, we constructed a suitable IONP, mito-magneto (MM), with solubility in DMF and insolubility in water. 44,45 MM also demonstrated a significantly high transverse relaxivity.…”

“…Inorganic NPs such as gold, quantum dots, and iron oxide can be used as contrast agents to image biological systems, employing their X-ray attenuation, fluorescence, and MRI signal enhancement properties. − Gold NPs or Cd-based quantum dots could be associated with heavy metal toxicity; iron oxide NP (IONP)-based contrast agents are among the safer options. − Most commercial IONPs with solubility in dimethylformamide (DMF) also demonstrate water solubility, and hence encapsulation and/or retention of these NPs in a hydrophobic nanocapsule is challenging. Thus, we constructed a suitable IONP, mito-magneto (MM), with solubility in DMF and insolubility in water. , MM also demonstrated a significantly high transverse relaxivity.…”

Understanding intracellular nanoparticle trafficking fates through spatiotemporally resolved magnetic nanoparticle recovery

“… 47,58,59 An example of this approach is found in many commercial products such as MACS separators. 60,61 Different magnet arrangements have been successfully exploited for the separation of magnetically tagged cells, 37,62 viruses, 63 DNA 64,65 and subcellular fractions from cell lysate such as mitochondria, 66 exosomes 67 and endocytic organelles. 29,68 …”