Abstract
Generating mammalian cells with specific mtDNA-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.
Article and author information
Author details
Funding
National Institutes of Health (T32CA009120)
- Alexander J Sercel
- Alexander N Patananan
National Institutes of Health (R21CA227480)
- Michael A Teitell
National Institutes of Health (P30CA016042)
- Michael A Teitell
CIRM (RT3-07678)
- Michael A Teitell
National Institutes of Health (T32GM007185)
- Alexander J Sercel
American Heart Association (18POST34080342)
- Alexander N Patananan
National Institutes of Health (T32GM008042)
- Amy K Yu
National Science Foundation (CBET 1404080)
- Pei-Yu Chiou
National Institutes of Health (R01GM114188)
- Pei-Yu Chiou
- Michael A Teitell
Air Force Office of Scientific Research (FA9550-15-1-0406)
- Pei-Yu Chiou
- Michael A Teitell
National Institutes of Health (R01GM073981)
- Michael A Teitell
National Institutes of Health (R01CA185189)
- Michael A Teitell
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Simon C Johnson, University of Washington, United States
Publication history
- Received: September 15, 2020
- Accepted: January 12, 2021
- Accepted Manuscript published: January 13, 2021 (version 1)
Copyright
© 2021, Sercel et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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