Figure 10.

The Knockout of the MMP3 significantly decreased the transmissive potential of tumoroid-derived EVs. EVs were collected after 6 days from the culture supernatants of tumoroids that formed in ULA plates. EVs were fluorescently labeled with BODIPY TR Ceramide (red). The labeled EVs or PBS were added to the MMP3-KO/palmG (green) tumoroids at a concentration of 5 μg/mL in the NCP-based 3D culture with the stemness-enhancing medium. The uptake of EVs was monitored over 24 h using the high contents screening (HCS) system. (A) Time-course imaging of EV uptake (red) by MMP3-KO/palmG tumoroids (green) for 24 h. Scale bar, 100 µm. (B) Red fluorescence intensities of transmitted EVs in MMP3-KO/palmG tumoroids. The average fluorescence intensity of the PBS treatment group at time point 0 h was evaluated as background and subtracted from raw values. n = 3, * p < 0.05, ** p < 0.01, and # p < 0.001 (LuM1-EVs versus MMP3-KO-EVs).