Fig. 2: Human tumor organoid culture and growth on microfluidic platform.

a On-platform growth of organoids: three separate patient-derived pancreatic ductal adenocarcinoma (PDAC) organoids in Matrigel from single cells to formation of differentiated morphology of complex 3D structures (scale bar 100 μm). b Organoids from two patients were grown in parallel in a traditional 24-well plate and on our microfluidic platform (scale bar 100 μm). After mature organoid formation, organoids were harvested, H&E stained, and their morphologies compared and analyzed. In both platforms, organoids from patient 1 exhibited back-to-back glands with a high degree of nuclear atypia and pleomorphism with an accumulation of apoptotic luminal necrotic cells. Organoids from patient 4 demonstrated a well-differentiated morphology with simple spherical organoids and uniform nuclear and cytoplasmic features with little or no accumulation of necrotic luminal cells. c Organoid growth curves of PDAC organoid samples derived from three patients grown from single cells for 7 days on the platform. Each dot represents the cross-sectional area of an individual organoid. Patient 1 (blue), Patient 2 (red), Patient 3 (green). d Long-term culture, growth, and fluorescent staining of fixed PDAC organoids on the platform. Nuclei staining (DAPI) and F-actin (Phalloidin) are demonstrated (scale bar 100 μm). e Multiple Z image slices or stacks of a group of fixed and fluorescently stained organoids with DAPI and phalloidin (scale bar 100 μm).