ADF basal medium: Advanced DMEM/F12 medium plus 1 mmol/L GlutaMAX, 1 mmol/L HEPES, and 100 U/mL penicillin/streptomycin.EN medium for tumor PDOs: ADF medium was supplemented with 10% Noggin conditioned medium (conditioned media were collected from cultures of HEK293 cells expressing recombinant Noggin proteins), 10 nmol/L gastrin I, 500 nmol/L A83–01, 10 mmol/L SB202190, 10 mmol/L nicotinamide, 1X B27 supplement, 1X N2 supplement, 1 mmol/L N-acetyl cysteine, and 50 ng/mL human recombinant EGF. The gene discussed is EGF; the disease is neoplasm.