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. Currently, 97 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

Assay Deteciton Regent Equipment
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

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


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

Title File format
Available F-PDO list Excel
F-PDO_summary PDF
Leaflet PDF
Poster_Evaluation of anticancer agents using patient derived tumor organoids PDF
Poster_Long Term culturable PDO Models F-PDO lines PDF
Poster_Characterization of F-PDO by 3D microscopic analysis PDF
Poster_F-PDO drug sensitivity and PDX Models from Japanese Patients PDF