We have developed the protocol of flow cytometry for characterizing diversity among oral stem-like cancer cells (SLCCs) using CD44, CD24, and aldehyde dehydrogenase (ALDH) in oral tumors. We are also reporting the protocol for tumor-derived explant cultures to develop oral cancer cell lines and enriching these diverse hybrid states of cancer cells in 3D spheroids from established cell lines. For complete details on the use and execution of this protocol, please refer to Vipparthi et al. (2022) and Vipparthi et al. (2021).
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