Pressure-Driven Mitochondrial Transfer Pipeline Generates Mammalian Cells of Desired Genetic Combinations and Fates

Alexander N Patananan; Alexander J Sercel; Ting-Hsiang Wu; Fasih M Ahsan; Alejandro Torres Jr; Stephanie A L Kennedy; Amy Vandiver; Amanda J Collier; Artin Mehrabi; Jon Van Lew; Lise Zakin; Noe Rodriguez; Marcos Sixto; Wael Tadros; Adam Lazar; Peter A Sieling; Thang L Nguyen; Emma R Dawson; Daniel Braas; Justin Golovato; Luis Cisneros; Charles Vaske; Kathrin Plath; Shahrooz Rabizadeh; Kayvan R Niazi; Pei-Yu Chiou; Michael A Teitell

doi:10.1016/j.celrep.2020.108562

Pressure-Driven Mitochondrial Transfer Pipeline Generates Mammalian Cells of Desired Genetic Combinations and Fates

Cell Rep. 2020 Dec 29;33(13):108562. doi: 10.1016/j.celrep.2020.108562.

Authors

Alexander N Patananan¹, Alexander J Sercel², Ting-Hsiang Wu³, Fasih M Ahsan¹, Alejandro Torres Jr¹, Stephanie A L Kennedy¹, Amy Vandiver⁴, Amanda J Collier⁵, Artin Mehrabi³, Jon Van Lew³, Lise Zakin⁶, Noe Rodriguez⁶, Marcos Sixto⁶, Wael Tadros⁶, Adam Lazar⁶, Peter A Sieling⁶, Thang L Nguyen⁷, Emma R Dawson¹, Daniel Braas⁸, Justin Golovato⁹, Luis Cisneros⁹, Charles Vaske⁹, Kathrin Plath⁵, Shahrooz Rabizadeh¹⁰, Kayvan R Niazi¹⁰, Pei-Yu Chiou¹¹, Michael A Teitell¹²

Affiliations

¹ Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
² Molecular Biology Interdepartmental Program, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ NanoCav LLC, Culver City, CA 90232, USA.
⁴ Division of Dermatology, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁵ Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA; Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁶ NantWorks, LLC, Culver City, CA 90232, USA.
⁷ Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁸ UCLA Metabolomics Center, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁹ NantOmics, LLC, Culver City, CA 90232, USA.
¹⁰ NanoCav LLC, Culver City, CA 90232, USA; NantWorks, LLC, Culver City, CA 90232, USA.
¹¹ Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Mechanical and Aerospace Engineering, University of California, Los Angeles, Los Angeles, CA 90095, USA; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
¹² Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA; Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA 90095, USA; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Pediatrics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA. Electronic address: mteitell@mednet.ucla.edu.

Abstract

Generating mammalian cells with desired mitochondrial DNA (mtDNA) sequences is enabling for studies of mitochondria, disease modeling, and potential regenerative therapies. MitoPunch, a high-throughput mitochondrial transfer device, produces cells with specific mtDNA-nuclear DNA (nDNA) combinations by transferring isolated mitochondria from mouse or human cells into primary or immortal mtDNA-deficient (ρ0) cells. Stable isolated mitochondrial recipient (SIMR) cells isolated in restrictive media permanently retain donor mtDNA and reacquire respiration. However, SIMR fibroblasts maintain a ρ0-like cell metabolome and transcriptome despite growth in restrictive media. We reprogrammed non-immortal SIMR fibroblasts into induced pluripotent stem cells (iPSCs) with subsequent differentiation into diverse functional cell types, including mesenchymal stem cells (MSCs), adipocytes, osteoblasts, and chondrocytes. Remarkably, after reprogramming and differentiation, SIMR fibroblasts molecularly and phenotypically resemble unmanipulated control fibroblasts carried through the same protocol. Thus, our MitoPunch "pipeline" enables the production of SIMR cells with unique mtDNA-nDNA combinations for additional studies and applications in multiple cell types.

Keywords: cell engineering; differentiation, MitoPunch, mitochondrial transplantation, mitochondrial replacement, mitonuclear communication, isolated mitochondria; mitochondrial transfer; mtDNA; reprogramming.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cell Differentiation
Cell Line
Cellular Reprogramming*
DNA, Mitochondrial / metabolism
Fibroblasts / metabolism*
Gene Transfer Techniques*
HEK293 Cells
High-Throughput Screening Assays / methods*
Humans
Induced Pluripotent Stem Cells / metabolism
Metabolome
Mice
Mice, Inbred C57BL
Mitochondria / genetics*
Mitochondria / metabolism*
Mitochondria / transplantation*
Transcriptome

Substances

DNA, Mitochondrial

Abstract

Publication types

MeSH terms

Substances

Grants and funding