An empirically refined workflow for generating patient-derived organoids from epithelial ovarian cancer
- 1Department of Obstetrics and Gynecology, Chung-Ang University Hospital
- 2Organoid Medical Research Center, Chung-Ang University
- 3Department of Microbiology and Molecular Genetics, University of California
Abstract
Patient-derived organoids (PDOs) can recapitulate selected features of original tumors and provide a useful system for studying epithelial ovarian cancer (EOC) in three-dimensional culture. Nonetheless, the protocol to generate EOC-PDOs has not yet been standardized. We therefore empirically refined the culture conditions and medium composition, considering the protocol effective when EOC PDOs were successfully established in five consecutive attempts. Tissue and ascites samples (from 29 and 8 patients, respectively) were used and the Syringe-extruded Organoid Basement membrane extract Assembly (SOBA) fragment culture method applied. Selected similarities in cell morphology, marker expression, and detectable alterations between organoids and patient-matched tumor tissue were assessed using pairwise comparisons. The refined workflow used a culture medium containing recombinant human R-Spondin 1 protein (250 ng/mL), Noggin, and NRG1, factors implicated in organoid maintenance and expansion. Ultimately, 31 EOC organoid lines successfully generated from 82.8% (24/29) of tissue and 87.5% (7/8) of ascites samples. The SOBA technique increased the 10 day PDO yield by 2.75-fold relative to the surface-attached dome method (p < 0.0001). This empirically refined workflow can be adjusted for specific experimental applications; however, further validation across larger and clinically diverse EOC cohorts is required.
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BibTeX
@article{ngo2026empirically,
title = {{An empirically refined workflow for generating patient-derived organoids from epithelial ovarian cancer}},
author = {Long Hoang Ngo and Que Thanh Thanh Nguyen and Hanh Thi Tuyet Vo and Mi-Kyung Kim and Sang-Kil Lee and Michael McClelland and Eun-Ju Lee},
journal = {Scientific Reports},
year = {2026},
month = jul,
doi = {10.1038/s41598-026-61960-2},
url = {https://doi.org/10.1038/s41598-026-61960-2},
note = {CC BY},
}
RIS
TY - JOUR
TI - An empirically refined workflow for generating patient-derived organoids from epithelial ovarian cancer
AU - Long Hoang Ngo
AU - Que Thanh Thanh Nguyen
AU - Hanh Thi Tuyet Vo
AU - Mi-Kyung Kim
AU - Sang-Kil Lee
AU - Michael McClelland
AU - Eun-Ju Lee
PY - 2026
DA - 2026/07/01
JO - Scientific Reports
DO - 10.1038/s41598-026-61960-2
UR - https://doi.org/10.1038/s41598-026-61960-2
AB - Patient-derived organoids (PDOs) can recapitulate selected features of original tumors and provide a useful system for studying epithelial ovarian cancer (EOC) in three-dimensional culture. Nonetheless, the protocol to generate EOC-PDOs has not yet been standardized. We therefore empirically refined the culture conditions and medium composition, considering the protocol effective when EOC PDOs were successfully established in five consecutive attempts. Tissue and ascites samples (from 29 and 8 patients, respectively) were used and the Syringe-extruded Organoid Basement membrane extract Assembly (SOBA) fragment culture method applied. Selected similarities in cell morphology, marker expression, and detectable alterations between organoids and patient-matched tumor tissue were assessed using pairwise comparisons. The refined workflow used a culture medium containing recombinant human R-Spondin 1 protein (250 ng/mL), Noggin, and NRG1, factors implicated in organoid maintenance and expansion. Ultimately, 31 EOC organoid lines successfully generated from 82.8% (24/29) of tissue and 87.5% (7/8) of ascites samples. The SOBA technique increased the 10 day PDO yield by 2.75-fold relative to the surface-attached dome method (p < 0.0001). This empirically refined workflow can be adjusted for specific experimental applications; however, further validation across larger and clinically diverse EOC cohorts is required.
SN - 2045-2322
ER -