TY - JOUR
T1 - Centrifugation-free magnetic isolation of functional mitochondria using paramagnetic iron oxide nanoparticles
AU - Banik, Bhabatosh
AU - Dhar, Shanta
N1 - Funding Information:
This protocol is based on our work that was partially carried out at the University of Georgia; the work was financially supported by the National Heart, Lung, and Blood Institute of the National Institutes of Health, under award number R56HL121392 to S.D. The protocol was written and submitted from the University of Miami. S.D. also thanks the American Heart Association for a National Scientist Award (14SDG18690009) and the Sylvester Comprehensive Cancer Center for currently supporting our work in this area.
PY - 2017/9/1
Y1 - 2017/9/1
N2 - Subcellular fractionation techniques are essential for cell biology and drug development studies. The emergence of organelle-targeted nanoparticle (NP) platforms necessitates the isolation of target organelles to study drug delivery and activity. Mitochondria-targeted NPs have attracted the attention of researchers around the globe, since mitochondrial dysfunctions can cause a wide range of diseases. Conventional mitochondria isolation methods involve high-speed centrifugation. The problem with high-speed centrifugation-based isolation of NP-loaded mitochondria is that NPs can pellet even if they are not bound to mitochondria. 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. Isolation of mitochondria using Mito-magneto eliminates artifacts typically associated with centrifugation-based isolation of NP-loaded mitochondria, thus producing intact, pure, and respiration-active mitochondria.
AB - Subcellular fractionation techniques are essential for cell biology and drug development studies. The emergence of organelle-targeted nanoparticle (NP) platforms necessitates the isolation of target organelles to study drug delivery and activity. Mitochondria-targeted NPs have attracted the attention of researchers around the globe, since mitochondrial dysfunctions can cause a wide range of diseases. Conventional mitochondria isolation methods involve high-speed centrifugation. The problem with high-speed centrifugation-based isolation of NP-loaded mitochondria is that NPs can pellet even if they are not bound to mitochondria. 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. Isolation of mitochondria using Mito-magneto eliminates artifacts typically associated with centrifugation-based isolation of NP-loaded mitochondria, thus producing intact, pure, and respiration-active mitochondria.
KW - Centrifugation-free method
KW - Iron oxide nanoparticles
KW - Magnetic resonance imaging
KW - Mitochondria isolation
UR - http://www.scopus.com/inward/record.url?scp=85030257493&partnerID=8YFLogxK
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U2 - 10.1002/cpcb.26
DO - 10.1002/cpcb.26
M3 - Article
C2 - 28862341
AN - SCOPUS:85030257493
VL - 2017
SP - 25.4.1-25.4.20
JO - Current Protocols in Cell Biology
JF - Current Protocols in Cell Biology
SN - 1934-2500
ER -