| International Session（Workshop）1(JGES・JSGE・JSGS・JSGCS) |
| Sat. November 23rd 9:30 - 12:00 Room 11: Portopia Hotel South Wing Topaz |
| Recombinant co-cultured pancreatic cancer tumoroids from EUS-FNB specimens | |||
| Christopher Jen Lock Khor1,2, Wai Leong Tam3, Damien Tan1,2 | |||
| 1Department of Gastroenterology and Hepatology, Singapore General Hospital, 2Duke-NUS Medical School, 3Genomic Institute of Singapore, A*Star | |||
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| Background: Pancreatic ductal adenocarcinoma (PDAC) tumoroids have been grown from surgically resected samples. Unfortunately, the majority of PDACs are unresectable. There is limited data on successful growth of tumoroids from EUS-guided fine needle biopsy (EUS-FNB) samples obtained at diagnosis. Tumoroid growth using current methods are limited by different growth factor requirements between tumor and stellate cells. Forming a tumoroid from separately grown tumor and stellate cells may improve its stability. Co-culturing the tumor and stellate tumoroids together may better recapitulate original tumor histology. Objective: 1. Assess the ability of EUS-FNB vs surgical samples to obtain sufficient tissue for tumoroid formation. 2. Form separate tumor and stellate cell lines followed by individual propagation into tumoroids. 3. Co-culture PDAC tumoroids from tumor and stellate cells to reconstitute a 3-dimensional tumoroid where histology can be recapitulated. Methods: PDAC cells were obtained via EUS-FNB or surgical resection at our institution. EUS-FNB was performed via a 22-gauge FNB needle through a standard curvilinear array echoendoscope. Tissue samples from surgical resection or FNB were digested with wash buffer and dispase solution to form a cell pellet. The cells were resuspended in cell media supplemented with growth factors. They were then seeded directly in tissue culture plates (to propagate tumor stellate cells) or on a layer of irradiated mouse feeder cells (to propagate tumor cells). Successful cell line establishment was defined as propagation of cells in culture media over at least 5 passages. Tumor and stellate cell lines were fluorescently-labelled. Tumor cell lines were seeded at 2500 cell density as monoculture. Tumor stellate cells were then added in a 1:1 ratio for co-culture. Success was defined by ability to grow in culture for at least 7 days. Results: Successful propagation of tumor cell lines occurred in 5 of 9 (55.5%) FNB samples and 11 of 20 (55.0%) surgical samples. Successful propagation of stellate cell lines occurred in 7 of 9 (77.8%) FNB samples and 12 of 20 (60.0%) surgical samples. Tumoroids were successfully grown from all 3 (100%) FNB samples and 6 of 7 (85.7%) surgical samples. Co-cultured tumoroids were successfully grown in all 6 surgical samples (100%). All 3 FNB samples are still undergoing co-culture. Recapitulation of histology with the original tumor from surgical samples were seen in all 6 (100%) tumoroids and 1 of 6 (16.7%) co-cultured tumoroid. Conclusion: PDAC tumoroids can be successfully grown from cells obtained via EUS-FNB at time of initial diagnosis, with similar success rates as surgical specimens. EUS FNB specimens are suitable for propagation of tumor and stellate cell lines, tumoroid culture and even co-culture with stellate cells to reconstitute the original tumor. |
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Index Term 1: pancreatic cancer Index Term 2: tumoroid |
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