Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, United States
Tianxing Man
Department of Mechanical and Aerospace Engineering, University of California, Los Angeles, Los Angeles, United States
Ting-Hsiang Wu
NanoCav, LLC, Culver City, United States; NantBio, Inc, and ImmunityBio, Inc, Culver City, United States
Amy K Yu
Molecular Biology Interdepartmental Doctoral Program, University of California, Los Angeles, Los Angeles, United States
Garret W Guyot
Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, United States
Shahrooz Rabizadeh
NanoCav, LLC, Culver City, United States; NantBio, Inc, and ImmunityBio, Inc, Culver City, United States; NantOmics, LLC, Culver City, United States; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, United States
Kayvan R Niazi
NanoCav, LLC, Culver City, United States; NantBio, Inc, and ImmunityBio, Inc, Culver City, United States; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, United States; Department of Bioengineering, University of California, Los Angeles, Los Angeles, United States
Pei-Yu Chiou
Department of Mechanical and Aerospace Engineering, University of California, Los Angeles, Los Angeles, United States; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, United States; Department of Bioengineering, University of California, Los Angeles, Los Angeles, United States
Molecular Biology Interdepartmental Doctoral Program, University of California, Los Angeles, Los Angeles, United States; Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, United States; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, United States; Department of Bioengineering, University of California, Los Angeles, Los Angeles, United States; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research University of California, Los Angeles, Los Angeles, United States; Department of Pediatrics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, United States; Jonsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, United States
Generating mammalian cells with specific mitochondrial DNA (mtDNA)–nuclear DNA (nDNA) combinations is desirable but difficult to achieve and would be enabling for studies of mitochondrial-nuclear communication and coordination in controlling cell fates and functions. We developed ‘MitoPunch’, a pressure-driven mitochondrial transfer device, to deliver isolated mitochondria into numerous target mammalian cells simultaneously. MitoPunch and MitoCeption, a previously described force-based mitochondrial transfer approach, both yield stable isolated mitochondrial recipient (SIMR) cells that permanently retain exogenous mtDNA, whereas coincubation of mitochondria with cells does not yield SIMR cells. Although a typical MitoPunch or MitoCeption delivery results in dozens of immortalized SIMR clones with restored oxidative phosphorylation, only MitoPunch can produce replication-limited, non-immortal human SIMR clones. The MitoPunch device is versatile, inexpensive to assemble, and easy to use for engineering mtDNA–nDNA combinations to enable fundamental studies and potential translational applications.
