Figure 7.

Troubleshooting: Pancreatic Tumor Organoids

(A) Representative bright-field images showing the result of culturing organoids with old media. Both panels show the growth of the same tumor organoid culture, that was either expanded for 8 days on 4-week-old medium (left panel), or expanded on freshly prepared medium (right panel). Scale bar, 500 μm.

(B) When freezing organoids, the organoids should have the correct size and not be too large. Representative bright-field microscopy images show the result of thawing a culture that was frozen when the organoids reached a large size. As can be seen, even after 11 days, no new organoids grew out from the thawed cells. This culture could not be recovered and was lost. Scale bar, 500 μm.

(C) Representative bright-field image of an example showing an organoid culture that recovered only poorly from thawing and was plated at a density that was too low (first panel) to result in good outgrowth of the culture (second panel). In this case, the culture could be saved by pooling the organoids that grew out at day 10 (second panel), passaging them and plating them in a decreased volume of BME/Matrigel. Scale bar, 500 μm.

(D) Representative bright-field image of an example showing an organoid culture derived from breast tissue that is attaching to the bottom of the culture plate. The surface where the 2D cells are growing is indicated with arrows. Scale bar, 500 μm.