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Protocol for tumor dissociation and fluorescence-activated cell sorting of human head and neck cancers.
Okolo, Ogoegbunam; Yu, Victoria; Flashner, Samuel; Martin, Cecilia; Nakagawa, Hiroshi; Lin, Derrick T; Puram, Sidharth V; Parikh, Anuraag S.
Affiliation
  • Okolo O; 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.
  • Yu V; 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.
  • Flashner S; Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA.
  • Martin C; 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.
  • Nakagawa H; 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,
  • Lin DT; 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.
  • Puram SV; 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. Electronic address: sidpuram@wustl.edu
  • Parikh AS; 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. Electronic address: asp2145@cumc.columbia.edu.
STAR Protoc ; 4(2): 102294, 2023 May 06.
Article in En | MEDLINE | ID: mdl-37149858
ABSTRACT
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.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Language: En Journal: STAR Protoc Year: 2023 Type: Article Affiliation country: United States
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Full text: 1 Collection: 01-internacional Database: MEDLINE Language: En Journal: STAR Protoc Year: 2023 Type: Article Affiliation country: United States