FIGURE 1.

PDOTS establishment and characterization scheme. Tumor spheroids with autologous immune cells were isolated after limited collagenase digestion of fresh tumor specimens. They were embedded in Matrigel and cultured within Human organoid medium composed of Ad‐DF+++ (Advanced DMEM/F12 supplemented with 2 mM Ultra glutamine, 10 mM HEPES, and 100/100 U/ml Penicillin/Streptomycin), 100 ng/ml Noggin, 0.1 μg/ml R‐spondin 1, 1x B27 supplement, 1.25 mM N‐Acetylcysteine, 10 mM nicotinamide, 50 ng/ml human recombinant EGF, 500 nM A83‐01, 3 μM SB202190, and 10 nM prostaglandin E2. PDOTS was passaged either trituration with pipet or dissociation with trypsin for 3 min at 37°C. Scale bar, 100 μm