Abstract 4872: 3D bioprinting of fresh NSCLC: The Lung3Dprint proof of concept study

Abstract 3D bioprinted models are emerging to bridge the gap between 2D systems and animal models, advancing the efficiency and accuracy of preclinical workflows. This work aimed to (i) develop an innovative methodology to generate patient-derived bioprinted lung cancer (PDBLC) models that aim to reflect the heterogeneity of the parent tumor, and (ii) evaluate these models as a relevant ex vivo drug screening tool. Fresh Non-Small Cell Lung Cancer (NSCLC) samples were obtained from March to October 2025 at Hospices Civils de Lyon, dissociated and embedded into a fibrinogen-alginate-gelatin bioink (0.5 to 5x106 cells/mL of bioink). PDBLC models were obtained by extrusion-based bioprinting using a Cellink BIO XTM, with post-conditioning polymerization using thrombin, transglutaminase, and calcium. PDBLC models were then placed in a dedicated lung cancer cell culture medium and analyzed at different time points, using a non-disruptive method (PrestoBlueTM assay) and disruptive methods (flow cytometry and immunohistochemistry). Flow cytometry markers include EpCAM and CK-7/-8 to identify tumor cells, CD31 for endothelial cells, CD45, CD3, and CD19 for leukocyte infiltrate, and FAP, ɑ-SMA and PDGFR-β for analysis of cancer-associated fibroblasts. Tumor and non-tumor cell contents were quantified as number of cells per object and their percentage of total live cells present. Nine samples have presently been bioprinted and analyzed. PrestoBlueTM assay enabled non-specific monitoring of the model’s metabolic activity over time. The median percentage of tumor cells in the bioink was estimated at 40% (range: 11 - 85). The percentage of viable tumor cells on days 7 and 14 were dependent on the initial concentration of cells in the bioink, ranging from 5 to 84%. The absolute number of tumor cells per PDBLC model tended to stabilize with time. The absolute percentage of viable tumor cells tended to increase between day 0 and day 14, with a median fold-change of 1.7-fold (range: -0.76 - 7.2). This was notably due to the decrease in the leukocyte infiltration as well as the endothelial subpopulation. Leukocyte subpopulation remaining 14 days after bioprinting was found to be T lymphocytes (CD45+ CD3+). Exposure of PDBLC models to conventional and targeted therapies used in NSCLC patients is ongoing and results will be presented. 3D bioprinting offers a relatively inexpensive and rapid means to evaluate fresh tumor samples. Our preliminary data show that our model not only supports fresh NSCLC cell survival for up to 22 days, but also maintains the primary tumor's heterogeneity over this period. The use of these 3D models as a drug-screening platform is being evaluated and would be relevant for translational research and clinical practice. Citation Format: Aurélie Cadiou, Clarisse Thiollier-Schmitt, Gabrielle Devret, Raphaële Guelminger, Tanguy Fenouil, Corinne Perrin, Camille Leonce, Samantha Ballesta, Gaëtan Pochon, Nicolas Alcala, Lars Petter Jordheim, Charles Dumontet, Michaël Duruisseaux. 3D bioprinting of fresh NSCLC: The Lung3Dprint proof of concept study abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 4872.