Isolation of slow-cycling cancer cells from lung patient-derived xenograft using carboxyfluorescein-succinimidyl ester retention-mediated cell sorting

• Published in: STAR protocols 2023-06, Vol.4 (2), p.102167-102167, Article 102167
• Main Authors: Cho, Jaebeom, Min, Hye-Young, Lee, Ho-Young
• Format: Article
• Language: English
• Subjects: Cancer; Cell Biology; Cell Isolation; Cell-based Assays; Flow Cytometry/Mass Cytometry; Health Sciences; Model Organisms; Molecular/Chemical Probes; Protocol
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2023.102167

The slow-cycling subpopulation plays an important role in anticancer drug resistance and tumor recurrence. Here, we describe a clinically relevant patient-derived xenograft model and a carboxyfluorescein succinimidyl ester dye that is diluted in a cell proliferation-dependent manner. We detail steps to separate active-cycling cancer cells and slow-cycling cancer cells (SCCs) in heterogeneous cancer populations to confirm their different cellular properties. This protocol can be used to distinguish SCCs, investigate their biology, and develop strategies for anticancer therapeutics. For complete details on the use and execution of this protocol, please refer to Cho et al. (2021).1 [Display omitted] •Reliable establishment of lung PDX mouse model•Efficient dissociation of PDX tumor and construction of PDX-derived primary cells•Isolation of slow-cycling cancer cells based on the level of CFSE dye retention•Proliferation level comparison among the CFSElow, CFSEmid, and CFSEhigh populations Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. The slow-cycling subpopulation plays an important role in anticancer drug resistance and tumor recurrence. Here, we describe a clinically relevant patient-derived xenograft model and a carboxyfluorescein succinimidyl ester dye that is diluted in a cell proliferation-dependent manner. We detail steps to separate active-cycling cancer cells and slow-cycling cancer cells (SCCs) in heterogeneous cancer populations to confirm their different cellular properties. This protocol can be used to distinguish SCCs, investigate their biology, and develop strategies for anticancer therapeutics.