Figure 2.
Platelet extravasation in the OvCa-Chip. (A) Time line describing the events and end points when endothelial cells, tumor cells, and platelets are introduced into the chip, followed by analysis of platelet extravasation. The tumor-endothelial coculture was established in the first 48 hours. Platelet perfusion was initiated at 48 hours, and the vascular alteration and subsequent platelet extravasation physiology was observed for up to 168 hours (7 days). Confocal micrographs showing 3D cross-sectional views of the OvCa-Chip consisting of cancer cells (blue, nuclei), vascular endothelial lumen (green, VE cadherin), and platelets (red, anti CD41) (B) and the Control-Chip consisting of control cells (blue), vascular endothelial lumen (green), and platelets (red) (C), every 24 hours after the platelets were introduced into the device. Bars represent 100 µm. (D) Confocal micrographs showing platelets (red) adhering to the vascular endothelium (Endo, green) and corresponding extravasation of platelets into the cancer cells (Cancer cells, blue) crossing the dysfunctional vascular barrier, at increasing time points when cancer cells were cocultured within OvCa-Chip. Bars represent 25 µm. (E) No prominent platelet adherence to the endothelial cells (Endo, green) and extravasation into control cells (Control cells, blue) was found in the Control-Chip. (F) After 72 hours, time-lapse images (brightfield and fluorescence overlaid; 10 minutes apart) taken within the tumor chamber showing platelets (red) extravasating from the luminal side (arrow) through the pore and entering into the cancer tissue compartment. Bar represents 25 µm. The quantitation of platelet adherence to the endothelium (G); extravasated platelet concentration observed within the tumor chamber (H) (●, OvCa-Chip; ▪, Control-Chip); and extravasated platelet concentration because of gain or loss of function of endothelial cells (I). Shaded area in panels G and H represents the 48-hour period before the introduction of platelets into the OvCa-Chip, when tumor-vessel coculture was established. Error bars indicate the mean ± SEM; n = 3 individual experiments. Two-tailed paired Student t test. *P < .05; **P < .01. ns, nonsignificant.

Platelet extravasation in the OvCa-Chip. (A) Time line describing the events and end points when endothelial cells, tumor cells, and platelets are introduced into the chip, followed by analysis of platelet extravasation. The tumor-endothelial coculture was established in the first 48 hours. Platelet perfusion was initiated at 48 hours, and the vascular alteration and subsequent platelet extravasation physiology was observed for up to 168 hours (7 days). Confocal micrographs showing 3D cross-sectional views of the OvCa-Chip consisting of cancer cells (blue, nuclei), vascular endothelial lumen (green, VE cadherin), and platelets (red, anti CD41) (B) and the Control-Chip consisting of control cells (blue), vascular endothelial lumen (green), and platelets (red) (C), every 24 hours after the platelets were introduced into the device. Bars represent 100 µm. (D) Confocal micrographs showing platelets (red) adhering to the vascular endothelium (Endo, green) and corresponding extravasation of platelets into the cancer cells (Cancer cells, blue) crossing the dysfunctional vascular barrier, at increasing time points when cancer cells were cocultured within OvCa-Chip. Bars represent 25 µm. (E) No prominent platelet adherence to the endothelial cells (Endo, green) and extravasation into control cells (Control cells, blue) was found in the Control-Chip. (F) After 72 hours, time-lapse images (brightfield and fluorescence overlaid; 10 minutes apart) taken within the tumor chamber showing platelets (red) extravasating from the luminal side (arrow) through the pore and entering into the cancer tissue compartment. Bar represents 25 µm. The quantitation of platelet adherence to the endothelium (G); extravasated platelet concentration observed within the tumor chamber (H) (●, OvCa-Chip; ▪, Control-Chip); and extravasated platelet concentration because of gain or loss of function of endothelial cells (I). Shaded area in panels G and H represents the 48-hour period before the introduction of platelets into the OvCa-Chip, when tumor-vessel coculture was established. Error bars indicate the mean ± SEM; n = 3 individual experiments. Two-tailed paired Student t test. *P < .05; **P < .01. ns, nonsignificant.

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