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© 2020 by John Wiley Sons, Inc. Basic Protocol 1 Generation of esophageal cancer PDO Basic Protocol 2 Propagation and cryopreservation of esophageal cancer PDO Basic Protocol 3 Imaged-based monitoring of organoid size and growth kinetics Basic Protocol 4 Harvesting esophageal cancer PDO for histological analyses Basic Protocol 5 PDO content analysis by flow cytometry Basic Protocol 6 Evaluation of drug response with determination of the half-inhibitory concentration (IC ) Support Protocol Production of RN in HEK293T cell con