Cryopreservation of Whole Tumor Biopsies from Rectal Cancer Patients Enable Phenotypic and In Vitro Functional Evaluation of Tumor-Infiltrating T Cells
Abstract
:Simple Summary
Abstract
1. Introduction
2. Results
2.1. The Cryopreservation Method Is Key for Flow Cytometric Analyses of Tumor Suspensions
2.2. Viable TILs Can Be Recovered from Cryopreserved Tumors for Multiparametric Phenotyping
2.3. Frequencies and Proportions of Major TIL Subsets Remain Stable in Cryopreserved Tumors
2.4. Tregs Are Reduced in Cryopreserved Tumors but Sufficient for Phenotypic Analyses
2.5. Concurrent Assessment of Unconventional T Cell Subsets in Cryopreserved Tumors
2.6. TILs Retain Inducible IFN-γ Responses after Cryopreservation
3. Discussion
4. Materials and Methods
4.1. Rectal Cancer Patients
4.2. Cryopreservation of Tumors
4.3. PBMC Isolation
4.4. Generation of Single Cell Suspensions from Fresh and Cryopreserved Tissues
4.5. Thawing of Cryopreserved Tissues
4.6. Flow Cytometry
4.7. In Vitro Stimulation
4.8. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Liang, F.; Rezapour, A.; Falk, P.; Angenete, E.; Yrlid, U. Cryopreservation of Whole Tumor Biopsies from Rectal Cancer Patients Enable Phenotypic and In Vitro Functional Evaluation of Tumor-Infiltrating T Cells. Cancers 2021, 13, 2428. https://doi.org/10.3390/cancers13102428
Liang F, Rezapour A, Falk P, Angenete E, Yrlid U. Cryopreservation of Whole Tumor Biopsies from Rectal Cancer Patients Enable Phenotypic and In Vitro Functional Evaluation of Tumor-Infiltrating T Cells. Cancers. 2021; 13(10):2428. https://doi.org/10.3390/cancers13102428
Chicago/Turabian StyleLiang, Frank, Azar Rezapour, Peter Falk, Eva Angenete, and Ulf Yrlid. 2021. "Cryopreservation of Whole Tumor Biopsies from Rectal Cancer Patients Enable Phenotypic and In Vitro Functional Evaluation of Tumor-Infiltrating T Cells" Cancers 13, no. 10: 2428. https://doi.org/10.3390/cancers13102428