Fig 1.
Size measurements of human normal dermal fibroblast (A), colon cancer (B), bladder cancer (C), and metastatic breast cancer (D) spheroids. Left panel: representative fluorescence microscopy images of a spheroid just after electroporation (8 pulses of 100 μs, 1000 V/cm, and 1 Hz) in buffer containing propidium iodide (PI) to visualize electropermeabilized cells and light microscopy images of another spheroid just before treatment (LM). Right panel: Growth curves of the spheroids after treatment with respectively 168 mM calcium (Ca), 1 mM bleomycin (Bleo), electroporation (EP), 168 mM calcium electroporation (Ca EP), electrochemotherapy using 1 mM bleomycin (Bleo EP), and untreated controls (Control). Measurements performed before treatment and at day 2, 3, and 4. Spheroid size is normalized to the size before treatment, means +/- SD, n = 5–6.
Fig 2.
Live/dead staining with Calcein-AM and EthD-1 of human normal dermal fibroblast, colon cancer, and bladder cancer spheroids 4 days after treatment with 168 mM calcium (Ca), 1 mM bleomycin (Bleo), electroporation (EP; 8 pulses of 100 μs, 1000 V/cm, and 1 Hz), 168 mM calcium electroporation (Ca EP), or electrochemotherapy using 1 mM bleomycin (Bleo EP), and of untreated controls (Control). Upper panels are calcein-AM staining (living cells), middle panels are EthD-1 staining (dead cells), and lower panels are merged images of living (green) and dead (red) cells.
Fig 3.
Intracellular ATP measurements of human normal dermal fibroblast, colon cancer, bladder cancer, and breast cancer spheroids 1, 4, 24, and 72 hours after treatment with 168 mM calcium (Ca), electroporation (EP; 8 pulses of 100 μs, 1000 V/cm, and 1 Hz), 168 mM calcium electroporation (Ca EP), high voltage EP (8 pulses of 100 μs, 5000 V/cm, and 1 Hz), and of untreated controls (Control). Means + SD, n = 3–5 (n = 2 for blank). Please note the difference in the y-axes.