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, 60 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)|
|Lung F-PDO (papillary adenocarcinoma)|
Assay systems using F-PDO
|Cell growth inhibition||ATP||CellTitier Glo (Promega)||Enspire (PerkinElmer)|
|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
|Annexin V/PI||Annexin V-FITC/PI (Nacalai)||FCM (SONY)|
|Immunostaining||Fluorescent immunostaining||Antibodies against Ki67, HER2, EGFR, actin, z0-1, and etc
|Confocal microscope and NoviSight（Olympus）|
|ADDC activity||Reporter gene expression||ADCC Bioassey Effector Cell
■ High-Throughput In Vitro Assay using Patient-Derived Tumor Organoids. Higa et al., J Vis Exp, 172, e62668 (2021)
■ Construction of in vitro patient‑derived tumor models to evaluate anticancer agents and cancer immunotherapy. Takahashi et al., ONCOLOGY LETTERS, 21, 406 (2021)
■ An in vitro system for evaluating molecular targeted drugs using lung patient-derived tumor organoids, Takahashi et al., Cells. 8 481 (2019).
■ Evaluation of anticancer agents using patient-derived tumor organoids characteristically similar to source tissues. Tamura et al., Oncology Reports ,
40: 635-646 (2018).
|Available F-PDO list||Excel|
|Leaflet_Long term-culturable patient-derived tumor organoid|
|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|