Stable transplantation of human mitochondrial DNA by high-throughput, pressurized isolated mitochondrial delivery

  1. Alexander J Sercel
  2. Alexander N Patananan
  3. Tianxing Man
  4. Ting-Hsiang Wu
  5. Amy K Yu
  6. Garret W Guyot
  7. Shahrooz Rabizadeh
  8. Kayvan R Niazi
  9. Pei-Yu Chiou
  10. Michael A Teitell  Is a corresponding author
  1. UCLA, United States
  2. NanoCav, LLC, NantBio, Inc, and ImmunityBio, Inc, United States
  3. 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

  1. Alexander J Sercel

    Molecular Biology Institute Interdepartmental Program, UCLA, Los Angeles, United States
    Competing interests
    No competing interests declared.
  2. Alexander N Patananan

    Pathology and Laboratory Medicine, UCLA, Los Angeles, United States
    Competing interests
    No competing interests declared.
  3. Tianxing Man

    Mechanical and Aerospace Engineering, UCLA, Los Angeles, United States
    Competing interests
    No competing interests declared.
  4. Ting-Hsiang Wu

    NanoCav, LLC, NantBio, Inc, and ImmunityBio, Inc, Culver City, United States
    Competing interests
    Ting-Hsiang Wu, T.-H.W. was an employee of NanoCav, LLC, and is currently employed by NantBio, Inc and ImmunityBio, Inc..
  5. Amy K Yu

    Molecular Biology Institute Interdepartmental Program, UCLA, Los Angeles, United States
    Competing interests
    No competing interests declared.
  6. Garret W Guyot

    Pathology and Laboratory Medicine, UCLA, Los Angeles, United States
    Competing interests
    No competing interests declared.
  7. Shahrooz Rabizadeh

    NanoCav, LLC, NantBio, Inc, and ImmunityBio, Inc, and NantOmics, LLC, Culver City, United States
    Competing interests
    Shahrooz Rabizadeh, S.R. is a board member of NanoCav, LLC, and employed by NantBio, Inc, ImmunityBio, Inc, and NantOmics, LLC..
  8. Kayvan R Niazi

    NanoCav, LLC, NantBio, Inc, and ImmunityBio, Inc, Culver City, United States
    Competing interests
    Kayvan R Niazi, K.R.N. is a board member of NanoCav, LLC, and employed by NantBio, Inc and ImmunityBio, Inc..
  9. Pei-Yu Chiou

    Mechanical and Aerospace Engineering, UCLA, Los Angeles, United States
    Competing interests
    Pei-Yu Chiou, P.-Y.C. is a co-founder, board member, shareholder, and consultant for NanoCav, LLC, a private start-up company working on mitochondrial transfer techniques and applications..
  10. Michael A Teitell

    Pathology and Laboratory Medicine, UCLA, Los Angeles, United States
    For correspondence
    mteitell@mednet.ucla.edu
    Competing interests
    Michael A Teitell, M.A.T. is a co-founder, board member, shareholder, and consultant for NanoCav, LLC, a private start-up company working on mitochondrial transfer techniques and applications..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4495-8750

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

  1. Simon C Johnson, University of Washington, United States

Version history

  1. Received: September 15, 2020
  2. Accepted: January 12, 2021
  3. Accepted Manuscript published: January 13, 2021 (version 1)
  4. Version of Record published: February 5, 2021 (version 2)

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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  1. Alexander J Sercel
  2. Alexander N Patananan
  3. Tianxing Man
  4. Ting-Hsiang Wu
  5. Amy K Yu
  6. Garret W Guyot
  7. Shahrooz Rabizadeh
  8. Kayvan R Niazi
  9. Pei-Yu Chiou
  10. Michael A Teitell
(2021)
Stable transplantation of human mitochondrial DNA by high-throughput, pressurized isolated mitochondrial delivery
eLife 10:e63102.
https://doi.org/10.7554/eLife.63102

Share this article

https://doi.org/10.7554/eLife.63102

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