Characterizing diversity among human oral stem-like cancer cells using flow cytometry

• Published in: STAR protocols 2022-12, Vol.3 (4), p.101763-101763, Article 101763
• Main Authors: Ghosh, Subhashis, Mitra, Paromita, Saha, Uday, Vipparthi, Kavya, Singh, Sandeep
• Format: Article
• Language: English
• Subjects: Aldehyde Dehydrogenase - metabolism; Cancer; Cell biology; Cell culture; Cell differentiation; Cell Line; Cell separation/fractionation; Flow Cytometry; Flow cytometry/Mass cytometry; Humans; Mouth Neoplasms; Protocol; Single cell; Stem cells
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101763

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). [Display omitted] •Enzymatic digestion protocol for surgically resected human oral tumor tissues•Flow-cytometry-based characterization of diversity in oral SLCCs•Derivation of primary cultures and cell lines from tumor explants•Protocol for growing 3D spheroid cultures of hybrid states of oral SLCCs Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.