Stable transplantation of human mitochondrial DNA by high-throughput, pressurized isolated mitochondrial delivery
- UCLA, United States
- NanoCav, LLC, NantBio, Inc, and ImmunityBio, Inc, United States
- NanoCav, LLC, NantBio, Inc, and ImmunityBio, Inc, and NantOmics, LLC, United States
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.
Data availability
Figure 1-source data 1. Numerical simulation of MitoPunch pressure generation during mitochondrial delivery. Cited in the legend of Figure 1.
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.
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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Stable transplantation of human mitochondrial DNA by high-throughput, pressurized isolated mitochondrial delivery
eLife 10:e63102.
https://doi.org/10.7554/eLife.63102
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