Portal de Periódicos da CAPES

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

2020; Cell Press; Volume: 33; Issue: 13 Linguagem: Inglês

10.1016/j.celrep.2020.108562

2639-1856

Alexander N. Patananan, Alexander J. Sercel, Ting‐Hsiang Wu, Fasih M. Ahsan, Alejandro Torres, Stephanie A.L. Kennedy, Amy R. Vandiver, Amanda J. Collier, Artin Mehrabi, Jon T. Van Lew, Lise Zakin, N. Andrade 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 Niazi, Pei‐Yu Chiou, Michael A. Teitell,

CRISPR and Genetic Engineering

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.