Long Term-culturable PDO Models
Patient-derived tumor organoids established in Fukushima project (F-PDOs), which can be cultured for over a half-year period, were established from clinical tumor tissues (e.g., lung, ovary, uterus, etc.). Data were acquired from histological analysis, whole-exome analysis, and comprehensive gene expression analysis of F-PDOs and their source tumor tissues. The comparative analysis proved that the characteristics of F-PDOs are similar to those of their source tumors. In addition, HLA typing of F-PDO is also performed. Currently, 62 F-PDO lines have been established (F-PDO list).
F-PDOs were used in experiments on cell growth inhibition by anticancer agents. A suitable high-throughput assay system, with 96- or 384-well plates, was designed for each F-PDO, and standard anticancer agents were evaluated. Notably, F-PDOs showed more resistance to these agents than conventional cancer cell lines. These results indicate that F-PDOs maintain the characteristics of their source tumors, which were resistant to these agents. The assay system using F-PDOs enables evaluation of anticancer agents under conditions that better reflect clinical conditions than conventional methods and help discover markers of pharmacological effects for anticancer agents. In addition, F-PDOs can be implanted into immunodeficient mice.
Lung F-PDO (squamous cell carcinoma) |
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Lung F-PDO (papillary adenocarcinoma) |
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F-PDO summary
Assay systems using F-PDO
Assay | Deteciton | Regent | Equipment |
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Cell growth inhibition | ATP | CellTitier Glo (Promega) | Enspire (PerkinElmer) |
Impedance | - | xCelligence (ACEA) | |
Cell area | - | IncuCyte (Essen) | |
Dehydrogenase activity | Cell Counting Kit-8 (Dojin) | Enspire (PerkinElmer) | |
Dead cell detection | Calcein | Calcein AM (Molecular Probes) | Enspire (PerkinElmer) |
Cyanine | IncuCyte Cytotox Green (Essen) | IncuCyte (Essen) | |
Lactate dehydrogenase activity | Cytotoxicity Detection Kit (Roche) | Enspire (PerkinElmer) | |
Apoptosis activity | Caspase-3/7 activity | ApoTox-Glo (Promega) | Enspire (PerkinElmer) |
Caspase-3/7 activity | CellEvent Caspase-3/7 Green (Molecular Probes) |
IncuCyte (Essen) | |
Annexin V/PI | Annexin V-FITC/PI (Nacalai) | FCM (SONY) | |
Immunostaining | Fluorescent immunostaining | Antibodies against Ki67, HER2, EGFR, actin, z0-1, and etc DAPI |
Confocal microscope and NoviSight(Olympus) |
ADDC activity | Reporter gene expression | ADCC Bioassey Effector Cell (BPS Bioscience) |
Enspire (PerkinElmer) |
Reference
■ High-Throughput In Vitro Assay using Patient-Derived Tumor Organoids. Higa et al., J Vis Exp, 172, e62668 (2021)
https://www.jove.com/t/62668/high-throughput-in-vitro-assay-using-patient-derived-tumor-organoids
■ Construction of in vitro patient‑derived tumor models to evaluate anticancer agents and cancer immunotherapy. Takahashi et al., ONCOLOGY LETTERS, 21, 406 (2021)
https://www.spandidos-publications.com/10.3892/ol.2021.12667
■ An in vitro system for evaluating molecular targeted drugs using lung patient-derived tumor organoids, Takahashi et al., Cells. 8 481 (2019).
https://www.mdpi.com/2073-4409/8/5/481
■ Evaluation of anticancer agents using patient-derived tumor organoids characteristically similar to source tissues. Tamura et al., Oncology Reports ,
40: 635-646 (2018).
https://www.spandidos-publications.com/or/40/2/635
Title | File format |
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Available F-PDO list | Excel |
F-PDO_summary | |
Leaflet_Long term-culturable patient-derived tumor organoid | |
Leaflet_immuno oncology | |
Poster_Evaluation of anticancer agents using patient derived tumor organoids | |
Poster_Long Term culturable PDO Models F-PDO lines | |
Poster_Characterization of F-PDO by 3D microscopic analysis | |
Poster_F-PDO drug sensitivity and PDX Models from Japanese Patients |