Background: Non-metastatic lymphadenopathy is challenging to diagnose. The comparative diagnostic performance of endobronchial ultrasound (EBUS)-guided transbronchial mediastinal cryobiopsy (TBMC) vs. EBUS-transbronchial needle aspiration (TBNA) remains debated. Methods: This multicenter randomized trial was conducted in three hospitals. Patients with at least one mediastinal and/or hilar lesion of >= 1 cm in the short axis who required diagnostic bronchoscopy were included. The patients were randomized in a 1:1 ratio to receive either EBUS-TBNA followed by EBUS-TBMC (EBUS-TBNA-first group) or EBUS-TBMC followed by EBUS-TBNA (EBUS-TBMC-first group). The primary outcome was the diagnostic yields of EBUS-TBMC and EBUS-TBNA. Findings: The overall diagnostic yield of EBUS-TBMC for non-metastatic lymphadenopathy was significantly higher than that of EBUS-TBNA for specific benign etiologies and lymphomas (97.1% vs. 79.9%, p < 0.001). In the subgroup analysis, EBUS-TBMC showed a higher sensitivity for sarcoidosis than EBUS-TBNA (98.0% vs. 82.7%, p < 0.001). All patients experienced grade 1 airway bleeding. Conclusions: EBUS-TBMC demonstrated a higher diagnostic yield than EBUS-TBNA for non-metastatic lymphadenopathy in a cohort almost exclusively composed of sarcoidosis cases, with a good safety profile. EBUS-TBMC is a potential first-line diagnostic tool for non-metastatic lymphadenopathy.