Oral squamous cell carcinoma (OSCC) is the most common form of head and neck cancer. The current standard for treating primary OSCC is surgical resection combined with radiotherapy and chemotherapy. Despite improved therapeutic strategies, OSCC has high rates of metastasis and mortality, with one in two patients dying of the disease. Patient-derived organoids have become promising cell culture systems for disease modeling and precision medicine. Here we describe the high-efficiency generation of organoids from OSCC patients, which can be maintained in the culture for the long term. We further provide protocols for characterizing OSCC organoids using histology and immunofluorescence staining.