Stable transplantation of human mitochondrial DNA by high-throughput, pressurized isolated mitochondrial delivery
Abstract
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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jan 13, 2021
- Source ID
- 10.7554/elife.63102
Entities
People
- Alexander J. Sercel
- Alexander N. Patananan
- Amy K Yu
- Garret W Guyot
- Kayvan Niazi
- Michael A Teitell
- Pei-yu Chiou
- Shahrooz Rabizadeh
- Tianxing Man
- Ting-hsiang Wu
Organizations
- Air Force Office of Scientific Research
- American Heart Association
- California Institute for Regenerative Medicine
- NantOmics
- National Institutes of Health
- National Science Foundation
- University of California
- University of California, Los Angeles