BACKGROUND:Establishing a diagnosis of pancreatic cystic lesions (PCLs) preoperatively still remains challenging. Recently, endoscopic ultrasound (EUS)-through-the-needle biopsy (EUS-TTNB) using microforceps in PCLs has been made available. AIM:To assess the efficacy and safety of EUS-TTNB in the diagnosis of PCLs. METHODS:We retrospectively collected data of patients with PCLs who underwent both EUS-fine-needle aspiration (FNA) for cytology and EUS-TTNB at our institution since 2016. EUS-FNA for cytology was followed by EUS-TTNB in the same session. Evaluation of the cyst location, primary diagnosis, adverse events, and comparison between the cytologic fluid analyses and histopathology was performed. Technical success of EUS-TTNB was defined as visible tissue present after biopsy. Clinical success was defined as the presence of a specimen adequate to make a histologic or cytologic diagnosis. RESULTS:A total of 56 patients (mean age 66.9 ± 11.7, 53.6% females) with PCLs were enrolled over the study period. The mean cyst size was 28.8 mm (12-85 mm). The EUS-TTNB procedure was technically successful in all patients (100%). The clinical success rate using EUS-TTNB was much higher than standard EUS-FNA, respectively 80.4% (45/56) vs 25% (14/56). Adverse events occurred in 2 patients (3.6%) who developed mild pancreatitis that resolved with medical therapy. Using TTNB specimens, 23 of 32 cases (71.9%) with intraductal papillary mucinous neoplasm were further differentiated into gastric type (19 patients) and pancreaticobiliary type (4 patients) based on immunochemical staining. CONCLUSION:EUS-TTNB for PCLs was technically feasible and had a favorable safety profile. Furthermore, the diagnostic yield for PCLs was much higher with EUS-TTNB than standard EUS-FNA cytology and fluid carcinoembryonic antigen. EUS-TTNB should be considered as an adjunct to EUS-FNA and cytologic analysis in the diagnosis and management of PCLs.