Establishment of organoids using residual samples from saline flushes during endoscopic ultrasound-guided fine-needle aspiration in patients with pancreatic cancer

Abstract

Background and study aims In patients with pancreatic cancer (PC), patient-derived organoid cultures can be useful tools for personalized drug selection and preclinical evaluation of novel therapies. To establish a less invasive method of creating organoids from a patient’s tumor, we examined whether PC organoids can be established using residual samples from saline flushes (RSSFs) during endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA). Methods Five patients with PC who underwent EUS-FNA were enrolled in a prospective study conducted at our institution. RSSFs obtained during EUS-FNA procedures were collected. An organoid culture was considered as established when ≥ 5 passages were successful. Organoid-derived xenografts were created using established organoids. Results EUS-FNA was performed using a 22- or 25-gauge lancet needle without complications. Patient-derived organoids were successfully established in four patients (80.0 %) with the complete medium and medium for the selection of KRAS mutants. Organoid-derived xenografts were successfully created and histologically similar to EUS-FNA samples. Conclusions Patient-derived PC organoids were successfully established using EUS-FNA RSSFs, which are produced as a byproduct of standard manipulations, but are usually not used for diagnosis. This method can be applied to all patients with PC, without additional invasive procedures, and can contribute to the development of personalized medicine and molecular research.

Simultaneous establishment of pancreatic cancer organoid and cancer‐associated fibroblast using a single‐pass endoscopic ultrasound‐guided fine‐needle biopsy specimen

“…Because approximately 80% of patients with pancreatic cancer are diagnosed in advanced stages, the only way to obtain tissue from these patients is endoscopic ultrasoundguided fine-needle biopsy (EUS-FNB). Establishing PDOs using EUS-FNB samples has been attempted recently with varying success rates, [8][9][10][11] and there has been no attempt to culture PDOs and CAFs simultaneously using EUS-FNB samples. Therefore, this study aimed to evaluate the feasibility of the simultaneous establishment of PDOs and CAFs using a single-pass FNB specimen from pancreatic cancer patients.…”

Oil blotting paper for formalin fixation increases endoscopic ultrasound‐guided tissue acquisition‐collected sample volumes on glass slides

“…Pathological sampling has recently been used for the definitive diagnosis and molecular testing for initiating targeted therapies. 38 , 39 Although EUS‐TA is the main sampling method for the diagnosis of solid pancreatic lesions, 8 , 40 , 41 , 42 samples obtained through EUS‐TA often do not meet the suitability criteria for CGP tests. 26 Moreover, while several studies have focused on optimal needle types and puncture methods to obtain adequate samples and on rapid and macroscopic on‐site evaluations, 15 , 43 , 44 , 45 , 46 , 47 , 48 studies that focus on the formalin fixation process are scarce.…”

“…Pathological sampling has recently been used for the definitive diagnosis and molecular testing for initiating targeted therapies 38,39 . Although EUS‐TA is the main sampling method for the diagnosis of solid pancreatic lesions, 8,40–42 samples obtained through EUS‐TA often do not meet the suitability criteria for CGP tests 26 .…”

Validation of a Novel EUS-FNB-Derived Organoid Co-Culture System for Drug Screening in Patients with Pancreatic Cancer

“…We also explored alternative methods of validation, namely if organoids could be classified as malignant or benign either by microscopic appearance of live organoids or presence of sustained growth in TSM, defined as the presence of viable organoids for five passages, a previously applied criterium for a "successful culture" in earlier publications [7,8,25]. While WTO cultures often had a cystic appearance with clear, thin-walled organoids (Figure 4A), they could also appear as solid, dense-looking organoids (Figure 4C).…”