A Three-Dimensional Culture Model of Reversibly Quiescent Myogenic Cells

Figure 2

Characterization of C2C12 myospheres. (a) Bar chart from cytofluorometer data showing the proportion of cells (±SD) in different phases of the cycle, in a monolayer and in myospheres of different ages. and . (b) Cytofluorometer plot comparing the cell cycle in cells proliferating in a monolayer and in 23-day myospheres. (c) Ki67 expression in a monolayer and 6-day myospheres. (d) Myosin expression evaluated by immunofluorescence. A labelled syncytium is at the periphery of a 9-day myosphere. (e) H&E staining of 9-day myospheres containing differentiating cells. Syncytia leaving the myospheres or internally located are indicated with (i), syncytia isolated in the medium after exit from a myosphere with (ii). (iii) indicates a mature myosphere that does not appear to contain syncytia. (e) in B and C, the symbol (iv) labels small myospheres dissolving after the detachment of syncytia. (f) Accumulation of extracellular matrix proteins in myospheres over time. Laminin expression in 3-day and 12-day myospheres. Scale bar: 50 μm in (c), (d), and (f) and 100 μm in (e).

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