Bidirectional transfer of mitochondria between cells. (A)
Visualization of mitochondrial transfer among chondrocytes (CH), between
CH and mesenchymal stromal cells (MSCs), and vice versa after 7 hours of
coculture. Donating cells were stained with MitoTracker (in red), CH
were stained with CellTrace (in blue), and F-actin of all cells was
stained with SiR-Actin (in green). Mitochondria transported between 2
cell types are indicated by white arrows. Scale bar = 25 µm. Time-lapse
videos can be found in the supplemental information. (B)
Transport of mitochondria through a tunneling nanotube between MSC and
CH. Donating MSCs were stained with DiD (in blue) and MitoTracker (in
red). F-actin of all cells was stained with SiR-Actin (in green). Image
taken after 16 hours of coculture. Scale bar = 25 µm. (C)
Flow cytometry analysis of (co)culture conditioned media (CM) for small
particles including mitochondria-containing microvesicles (in red).
Noise and particles negative for both dyes are depicted in orange, and
cells (upper gate) are depicted in gray. Intensity of MitoTracker
(561/610 nm) is depicted on the x axis, and intensity
of CellTrace (405/450 nm) is depicted on the y axis.
MSC− and CH− are unstained. MSC+ and CH+ are dual-stained for
MitoTracker and CellTrace. In unidirectional cocultures (lower panels,
left and middle), the first cell type is dual-stained, while the other
is unstained. In the bidirectional coculture (lower panel, right), both
cell types are dual-stained. In all cases, 10,000 events were
recorded.