Pressure-Driven Mitochondrial Transfer Pipeline Generates Mammalian Cells of Desired Genetic Combinations and Fates
| Autor: | Thang L. Nguyen, Sixto Marcos, Pei-Yu Chiou, Shahrooz Rabizadeh, Alexander N. Patananan, Adam Lazar, Jon T. Van Lew, Noe Rodriguez, Luis Cisneros, Peter A. Sieling, Daniel Braas, Fasih M. Ahsan, Charles J. Vaske, Michael A. Teitell, Amy R. Vandiver, Amanda J. Collier, Artin Mehrabi, Alexander J. Sercel, Stephanie A.L. Kennedy, Lise Zakin, Emma R. Dawson, Tadros Wael, Alejandro Torres, Kayvan Niazi, Ting-Hsiang Wu, Justin Golovato, Kathrin Plath |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Cell Medical Physiology Mitochondrion Inbred C57BL isolated mitochondria Regenerative Medicine Transcriptome Mice 0302 clinical medicine Stem Cell Research - Nonembryonic - Human Induced pluripotent stem cell mtDNA Gene Transfer Techniques mitochondrial transfer Cell Differentiation differentiation Cellular Reprogramming Cell biology Mitochondria Mitochondrial medicine.anatomical_structure mitonuclear communication Metabolome mitochondrial transplantation Reprogramming Mitochondrial DNA Cell type 1.1 Normal biological development and functioning Induced Pluripotent Stem Cells Biology cell engineering DNA Mitochondrial General Biochemistry Genetics and Molecular Biology Article Cell Line 03 medical and health sciences Underpinning research Clinical Research medicine Genetics Animals Humans Stem Cell Research - Induced Pluripotent Stem Cell Mesenchymal stem cell reprogramming DNA Fibroblasts Stem Cell Research High-Throughput Screening Assays Mice Inbred C57BL MitoPunch 030104 developmental biology HEK293 Cells mitochondrial replacement Generic health relevance Biochemistry and Cell Biology 030217 neurology & neurosurgery |
| Zdroj: | Cell reports, vol 33, iss 13 Cell reports |
| Popis: | SUMMARY 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. Graphical Abstract In Brief Patananan and colleagues demonstrate a pipeline for transferring isolated mitochondria into mtDNA-deficient recipient cells. mtDNA-depleted fibroblasts permanently retain acquired non-native mtDNA through cell fate transitions. Initially, mitochondrial recipients show mtDNA-deficient cell transcriptome and metabolome profiles, with improvement to control profiles by reprogramming to pluripotency and subsequent differentiation. |
| Databáze: | OpenAIRE |
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