Abstract:

Objective: To explore the feasibility of employing vitrification cryopreservation, an advanced cryopreservation technique, for preserving surgically resected lung cancer tissues to facilitate subsequent organoid generation. Methods: We prospectively collected tumor specimens from 12 patients with histologically confirmed lung cancer at Beijing Chest Hospital, Capital Medical University from July to August 2024. Following standardized pathological procedures, the tissues were immediately immersed in preservation medium, maintained at 4 ℃ during transportation, and processed within 2 hours. Each specimen was systematically divided into three aliquots of approximately 0.5 cm³. One was processed fresh for organoid culture (serving as the control), while the remaining two were subjected to vitrification cryopreservation and stored in a vapor-phase liquid nitrogen tank. The cryopreserved samples were subsequently thawed at two time points (1 week and 4 months) for comparative assessment of organoid culture. Results: All fresh specimens (100.0%, 12/12) successfully generated organoids. Vitrified samples showed success rates of 75.0% (9/12) and 50.0% (6/12) at 1-week and 4-month time points, respectively. Crucially, cryopreserved-derived organoids maintained morphological characteristics and tumor marker expression profiles comparable to those of the fresh controls. Notably, all specimens with post-thaw viability <55% after 4 months of storage failed to organoid generation. Conclusion: This small-scale study indicates that although long-term vitrification cryopreservation exerts certain detrimental effects on cell viability, the processed surgical specimens can still meet the requirements for organoid construction.

Key words: Lung neoplasms, Freezing, Specimen handling, Organoids