The invention relates to the field of culture of tumor organoids, and in particular relates to a special medium used for in-vitro culture of esophagus cancer tumor organoid based on human esophagus cancer tissue and a culture method. The special medium comprises a receptor tyrosine kinase ligand, an Rho-associated coiled-coil protein kinase inhibitor, vitamin and hormone, an antioxidant, and an agonist for inhibiting cell differentiation pathway. According to the method, through the step-by-step culture technology adopting conditional dedifferentiation, tumor cells rapidly amplify within a short time to form the organoid. Through the step-by-step culture technology, primary cells rapidly form the organoid, meanwhile, adequate cells are formed within the effective time, so that the follow-up experiment operation can be conveniently carried out, and therefore, the culture speed and success rate of the organoid are improved.