Transferring isolated mitochondria into tissue culture cells

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Transferring isolated mitochondria into tissue culture cells

We have developed a new method for introducing large numbers of isolated mitochondria into tissue culture cells. Direct microinjection of mitochondria into typical mammalian cells has been found to be impractical due to the large size of mitochondria relative to microinjection needles. To circumvent this problem, we inject isolated mitochondria through appropriately sized microinjection needles...

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Experiments were undertaken to study the effect of introducing isolated mammalian cell mito chondria into tissue culture cells. The D N A of isolated mitochondria was labeled in vitro with 3 Hthymidine. Incorporation of 3H-thymidine into mitochondrial D N A was increased tenfold by the addition of bovine serum albumin and sucrose to the assay. Labeled HeLa cell mitochondria were fused with WI38...

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Isolation of mitochondria from tissue culture cells.

The number of mitochondria per cell varies substantially from cell line to cell line. For example, human HeLa cells contain at least twice as many mitochondria as smaller mouse L cells. This protocol starts with a washed cell pellet of 1-2 mL derived from ∼10⁹ cells grown in culture. The cells are swollen in a hypotonic buffer and ruptured with a Dounce or Potter-Elvehjem homogenizer using a ti...

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ژورنال

عنوان ژورنال: Nucleic Acids Research

سال: 2012

ISSN: 1362-4962,0305-1048

DOI: 10.1093/nar/gks639