Chapter title |
Testing Susceptibility of Patient-Derived Organoid Cultures to Therapies: Pharmacotyping
|
---|---|
Book title |
Phenotypic Screening
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7847-2_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7846-5, 978-1-4939-7847-2
|
Authors |
Richard A. Burkhart, Lindsey A. Baker, Hervé Tiriac |
Abstract |
Increasingly, patient models of disease are being utilized to facilitate precision medicine approaches through molecular characterization or direct chemotherapeutic testing. Organoids, 3-dimensional (3D) cultures of neoplastic cells derived from primary tumor specimens, represent an ideal platform for these types of studies because benchtop protocols previously developed for 2-dimensional cell lines can be adapted for use. These protocols include directly testing the survival of these organoid cultures when exposed to clinically relevant chemotherapeutic agents, a process we have called pharmacotyping. In this protocol, established tumor-derived organoid cultures are dissociated into single cells, plated in a 3D gel matrix, and exposed to pharmacologic agents. While our protocol has been developed for use with patient-derived pancreatic ductal adenocarcinoma organoids, with minor modifications to the dissociation and medium conditions, this protocol could be adapted for use with a wide range of organoid cultures. We further describe our standard ATP-based assay to determine cellular survival. This protocol can be scaled for use in high-throughput assays. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Italy | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 22 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Master | 4 | 18% |
Student > Bachelor | 3 | 14% |
Researcher | 3 | 14% |
Student > Ph. D. Student | 2 | 9% |
Lecturer > Senior Lecturer | 1 | 5% |
Other | 2 | 9% |
Unknown | 7 | 32% |
Readers by discipline | Count | As % |
---|---|---|
Medicine and Dentistry | 6 | 27% |
Biochemistry, Genetics and Molecular Biology | 4 | 18% |
Pharmacology, Toxicology and Pharmaceutical Science | 2 | 9% |
Agricultural and Biological Sciences | 2 | 9% |
Chemistry | 2 | 9% |
Other | 0 | 0% |
Unknown | 6 | 27% |