Protocol for tumor dissociation and fluorescence-activated cell sorting of human head and neck cancers

Ogoegbunam Okolo; Victoria Yu; Samuel Flashner; Cecilia Martin; Hiroshi Nakagawa; Derrick T. Lin; Sidharth V. Puram; Anuraag S. Parikh

STAR Protocols (Jun 2023)

Protocol for tumor dissociation and fluorescence-activated cell sorting of human head and neck cancers

Ogoegbunam Okolo,
Victoria Yu,
Samuel Flashner,
Cecilia Martin,
Hiroshi Nakagawa,
Derrick T. Lin,
Sidharth V. Puram,
Anuraag S. Parikh

Affiliations

Ogoegbunam Okolo: Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA; Columbia Vagelos College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA
Victoria Yu: Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA; Department of Otolaryngology-Head and Neck Surgery, Columbia University, New York, NY 10032, USA
Samuel Flashner: Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA
Cecilia Martin: Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA; Organoid and Cell Culture Core, Columbia University Digestive and Liver Diseases Research Center, Columbia University, New York, NY 10032, USA
Hiroshi Nakagawa: Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA; Organoid and Cell Culture Core, Columbia University Digestive and Liver Diseases Research Center, Columbia University, New York, NY 10032, USA; Division of Digestive and Liver Diseases, Department of Medicine, Columbia University, New York, NY 10032, USA
Derrick T. Lin: Department of Otolaryngology, Massachusetts Eye and Ear, Harvard University, Boston, MA 02114, USA; Department of Otolaryngology, Harvard Medical School, Harvard University, Boston, MA 02114, USA
Sidharth V. Puram: Department of Otolaryngology, Washington University School of Medicine, Washington University in St. Louis, St. Louis, MO 63110, USA; Department of Genetics, Washington University School of Medicine, Washington University in St. Louis, St. Louis, MO 63110, USA; Corresponding author
Anuraag S. Parikh: Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA; Department of Otolaryngology-Head and Neck Surgery, Columbia University, New York, NY 10032, USA; Corresponding author

Journal volume & issue: Vol. 4, no. 2
p. 102294

Abstract

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Summary: Tumors originating from the head and neck represent diverse histologies and are comprised of several cell types, including malignant cells, cancer-associated fibroblasts, endothelial cells, and immune cells. In this protocol, we describe a step-by-step approach for the dissociation of fresh human head and neck tumor specimens, followed by isolation of viable single cells using fluorescence-activated cell sorting. Our protocol facilitates the effective downstream use of techniques, including single-cell RNA sequencing and generation of three-dimensional patient-derived organoids.For complete details on the use and execution of this protocol, please refer to Puram et al. (2017)1 and Parikh et al. (2022).2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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