Generation and Characterization of Patient-Derived Head and Neck, Oral, and Esophageal Cancer Organoids.

Autor: Karakasheva TA; Division of Gastroenterology, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.; Epithelial Biology Center, Division of Pediatric Gastroenterology, Hepatology, and Nutrition, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania., Kijima T; Division of Digestive and Liver Diseases, Department of Medicine and Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York., Shimonosono M; Division of Digestive and Liver Diseases, Department of Medicine and Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York., Maekawa H; Division of Digestive and Liver Diseases, Department of Medicine and Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York., Sahu V; Division of Digestive and Liver Diseases, Department of Medicine and Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York., Gabre JT; Division of Gastroenterology, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania., Cruz-Acuña R; Division of Gastroenterology, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania., Giroux V; Department of Immunology and Cell Biology, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec, Canada., Sangwan V; Department of Surgery, Montreal General Hospital, McGill University, Montreal, Quebec, Canada., Whelan KA; Fels Institute for Cancer Research and Molecular Biology, Department of Pathology and Molecular Medicine, Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania., Natsugoe S; Department of Digestive Surgery, Breast and Thyroid Surgery, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan., Yoon AJ; Division of Oral & Maxillofacial Pathology and Department of Pathology & Cell Biology, Columbia University Irving Medical Center, New York, New York., Philipone E; Division of Oral & Maxillofacial Pathology and Department of Pathology & Cell Biology, Columbia University Irving Medical Center, New York, New York., Klein-Szanto AJ; Histopathology Facility, Fox Chase Cancer Center, Philadelphia, Pennsylvania., Ginsberg GG; Division of Gastroenterology, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania., Falk GW; Division of Gastroenterology, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania., Abrams JA; Division of Digestive and Liver Diseases, Department of Medicine and Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York., Que J; Division of Digestive and Liver Diseases, Department of Medicine and Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York., Basu D; Department of Otorhinolaryngology, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania., Ferri L; Department of Immunology and Cell Biology, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec, Canada., Diehl JA; Department of Biochemistry, School of Medicine and Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, Ohio., Bass AJ; Dana-Farber Cancer Institute, Harvard Medical School, Broad Institute, Boston, Massachusetts., Wang TC; Division of Digestive and Liver Diseases, Department of Medicine and Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York., Rustgi AK; Division of Digestive and Liver Diseases, Department of Medicine and Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York., Nakagawa H; Division of Digestive and Liver Diseases, Department of Medicine and Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York.
Jazyk: angličtina
Zdroj: Current protocols in stem cell biology [Curr Protoc Stem Cell Biol] 2020 Jun; Vol. 53 (1), pp. e109.
DOI: 10.1002/cpsc.109
Abstrakt: Esophageal cancers comprise adenocarcinoma and squamous cell carcinoma, two distinct histologic subtypes. Both are difficult to treat and among the deadliest human malignancies. We describe protocols to initiate, grow, passage, and characterize patient-derived organoids (PDO) of esophageal cancers, as well as squamous cell carcinomas of oral/head-and-neck and anal origin. Formed rapidly (<14 days) from a single-cell suspension embedded in basement membrane matrix, esophageal cancer PDO recapitulate the histology of the original tumors. Additionally, we provide guidelines for morphological analyses and drug testing coupled with functional assessment of cell response to conventional chemotherapeutics and other pharmacological agents in concert with emerging automated imaging platforms. Predicting drug sensitivity and potential therapy resistance mechanisms in a moderate-to-high throughput manner, esophageal cancer PDO are highly translatable in personalized medicine for customized esophageal cancer treatments. © 2020 by John Wiley & Sons, Inc. Basic Protocol 1: Generation of esophageal cancer PDO Basic Protocol 2: Propagation and cryopreservation of esophageal cancer PDO Basic Protocol 3: Imaged-based monitoring of organoid size and growth kinetics Basic Protocol 4: Harvesting esophageal cancer PDO for histological analyses Basic Protocol 5: PDO content analysis by flow cytometry Basic Protocol 6: Evaluation of drug response with determination of the half-inhibitory concentration (IC 50 ) Support Protocol: Production of RN in HEK293T cell conditioned medium.
(© 2020 John Wiley & Sons, Inc.)
Databáze: MEDLINE
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