Journal
BIOLOGY OPEN
Volume 6, Issue 12, Pages 1960-1965Publisher
COMPANY OF BIOLOGISTS LTD
DOI: 10.1242/bio.024869
Keywords
Cell fusion; Mitochondrial cloning; Homoplasmic mutation of mtDNA
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Funding
- Japan Society for the Promotion of Science [22220009, 26650068, 16K07207]
- Center of Innovation Program from Japan Science and Technology Agency
- Single Cell Project from RIKEN
- Grants-in-Aid for Scientific Research [26650068, 16K07207] Funding Source: KAKEN
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Quantitative control of mitochondria transfer between live cells is a promising approach for genetic manipulation of mitochondrial DNA (mtDNA) because single mitochondrion transfer to a mtDNA-less (rho(0)) cell potentially leads to homoplasmy of mtDNA. In this paper, we describe a method for quantitative control of mitochondria transfer between live single cells. For this purpose, we fabricated novel microfluidic devices having cell paring structures with a 4.1, 5.6 or 10.0 mu m-length microtunnel. When cells were fused through a microtunnel using the Sendai virus envelope-based method, a strictured cytoplasmic connection was achieved with a length corresponding to that of the microtunnel. Elongation of the cytoplasmic connection led to a decrease in mitochondria transfer to the fusion partner. Moreover, some cell pairs that fused through a 10.0 mu m length microtunnel showed single mitochondrion transfer. Fused cells were spontaneously disconnected from each other when they were recovered in a normal culture medium. These results suggest that our cell fusion method can perform quantitative control of mitochondria transfer that includes a single mitochondrion transfer.
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