Generation and characterization of patient-derived esophageal cancer organoids

Esophageal cancers comprise adenocarcinoma and squamous cell carcinoma, two distinct histologic subtypes. Both are difficult to treat and amongst the deadliest human malignancies. We describe protocols to initiate, grow, passage and characterize patient-derived organoids (PDO) of esophageal cancers as well as squamous cell carcinomas of oral/head-and-neck and anal origin. Formed rapidly (< 14 days) from a single cell suspension embedded in basement membrane matrix, esophageal cancer PDO recapitulate the histology of the original tumors. Additionally, we provide guidelines for morphological analyses and drug testing coupled with functional assessment of cell response to conventional chemotherapeutics and other pharmacological agents in concert with emerging automated imaging platforms. Predicting drug sensitivity and potential therapy resistance mechanisms in a moderate-to-high throughput manner, esophageal cancer PDO are highly translatable in personalized medicine for customized esophageal cancer treatments.