Generation and characterization of 3D bioprinted human endometrial cancer models

Endometrial carcinoma (EC) is among the most common gynaecological malignancies, with increasing incidence and limited therapeutic options in advanced and recurrent disease. The molecular mechanisms underlying tumour progression and drug resistance are not fully understood, highlighting the need for reliable and biologically relevant preclinical models. Conventional two-dimensional (2D) cell cultures fail to reproduce the complex three-dimensional (3D) architecture of tumours and the dynamic interactions between cancer cells and the extracellular matrix. In contrast, 3D culture systems more faithfully mimic tumour microenvironment, thereby offering enhanced biological relevance for the study of cancer pathophysiology and therapeutic resistance. The study aimed to develop and characterize 3D EC models, including spheroids and bioprinted constructs encapsulated in naturally derived bioinks, to establish more predictive platforms for drug evaluation compared to conventional 2D cultures. Three EC cell lines were employed to capture tumour heterogeneity. Spheroids were generated at different cell densities and analyzed for morphology, aggregation capacity and viability. The 3D bioprinted models were assessed for morphology, viability and molecular features. Gene and miRNA expression analyses revealed modulation of pathway associated with extracellular matrix remodelling, proliferation, migration and chemoresistance in 3D compared to 2D systems. Pharmacological testing with doxorubicin, paclitaxel, cisplatin and carboplatin revealed reduced cytotoxic efficacy in 3D models. Importantly, 3D bioprinted models enabled the evaluation of long-term drug effects and adaptive response. Overall, the 3D bioprinted EC models exhibit biologically and pharmacologically relevant features that more closely resemble in-vivo tumour behaviour, representing valuable tools for investigating EC progression and improving the predictive power of preclinical drug testing.