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. Author manuscript; available in PMC: 2016 Jun 26.
Published in final edited form as: Oncogene. 2013 Nov 4;33(45):5238–5250. doi: 10.1038/onc.2013.467

Figure 3.

Figure 3

Induction of cancer stem cells with ‘self-renewal’ capacity by reducing mtDNA copy number. (a and e) Number of spheres formed by parental, mtDNA-reduced and reverted MCF10A (a) and MCF7 (e) cells when grown on low attachment surface. The primary spheres (Gen1) were disintegrated by pipetting and replated further, and secondary spheres (Gen 2) formed by the parental, mtDNA-reduced and reverted cells were counted. (b) Bright-field images of mammospheres formed after 14 days in 3D culture (2% matrigel) by parental, mtDNA-reduced and reverted MCF10A and MCF10A cells. Scale bar, 10 μm. (c, h) CD44 and CD24 expression levels in MCF10A (c) and MCF7 (h) parental, mtDNA-reduced and reverted cells quantified by flow cytometry analysis. (d, i) Immunofluorescence images (× 40 magnification) showing expression levels of CD44 and CD24 in parental, mtDNA-reduced and reverted mtDNA-MCF10A (d) and MCF7 (i) cells. Scale bar, 20 μm. (f) Bright-field images of spheres formed by parental, mtDNA-reduced and reverted MCF7 cells grown on low attachment surface for two generations. Parental and reverted cells lose their sphere-forming capacity in Gen2 and grow as monolayers. Scale bar, 10 μm. (g) Hematoxylin and eosin staining of the spheres formed by parental and mtDNA-reduced MCF7 cells as described in f above. Scale bar, 10 μm.