Journal: Cell reports
Article Title: Carcinoma-associated mesenchymal stem cells promote ovarian cancer heterogeneity and metastasis through mitochondrial transfer
doi: 10.1016/j.celrep.2024.114551
Figure Lengend Snippet: (A) Representative images from coculture demonstrating direct CA-MSC to TC mitochondrial transfer. CA-MSC mitochondria labeled with lentiviral COX8-GFP (green), while endogenous TC mitochondria are COX8-dsRed (orange). (i) Maximum projection image of representative coculture z stack. scale bar, 50 μm. (ii) Three-dimensional (3D) rendering of a subset of (i) showing TC +mitoTrans cell and (iii) representative z slices. (B) Transfer of GFP-CA-MSC mitochondria to TCs quantified using flow cytometry. Six individual patient (pt)-derived CA-MSCs are represented (i) and three individual patient-derived CA-MSCs donating to three individual ovarian cancer cell lines are represented (ii). (C) CA-MSC to TC mitochondrial transfer under indirect coculture vs. direct coculture ( n = 5 individual experiments). (D) Comparison of CA-MSCs’ and normal MSCs’ (derived from patients without cancer) ability to transfer mitochondria to TCs ( n = 3 individual experiments). (E) Change in CA-MSC to TC mitochondrial transfer under adherent vs. non-adherent conditions ( n = 5 individual experiments). (F) Change in CA-MSC to TC mitochondrial transfer under normoxic (21% O 2 ) vs. hypoxic (1% O 2 ) conditions ( n = 3 individual experiments). (G) CA-MSC mitochondrial transfer quantified comparing donation to TCs with the highest active mitochondrial content (top 20%, mito rich) and lowest active mitochondrial content (lower 20%, mito poor), as assessed by MitoTracker Deep Red staining. In (B)–(D), CA-MSCs were cocultured, resulting in a 10:1 TC:CA-MSC ratio. * p < 0.05. Data are presented as the mean ± SEM; displayed p values obtained using Student’s t test.
Article Snippet: shRNA constructs targeting MIRO1/RHOT1 (HSH070029-LVRU6MP, GeneCopoeia, Rockville, MD) were packaged into lentiviral particles using a 3rd Generation Lentiviral Packaging Kit (Applied Biological Materials Inc., Richmond, BC).
Techniques: Labeling, Flow Cytometry, Derivative Assay, Comparison, Staining
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