Comparison of advanced in vitro models for toxicology assessment.
| Model type | Physiological relevance(ECM mimicry, cell-cell interactions, gradients, tissue architecture) | Throughput capability(HTS compatibility) | Suitability for immunotoxicology(immune cell integration) | Suitability for carcinogenicity(TME mimicry, drug screening) | Key advantages | Key challenges |
|---|---|---|---|---|---|---|
| Spheroids | Moderate: 3D cell-cell interactions, gradients, basic TME mimicry [37, 38]. | High: especially when using established cell lines [31, 36]. | Good: heterotypic spheroids can be co-cultured with immune cells [35, 52]. | Good: models drug resistance, TME mimicry, and proliferation effectively in a 3D context [37, 46]. | Simplicity, cost-effectiveness, scalability, HTS compatibility [31, 36]. | Lack of complex tissue architecture, limited long-term stability for primary cells, batch-to-batch size variability [8, 35]. |
| Organoids | High: self-organization creates organ-specific cell types, complex tissue architecture, and patient heterogeneity [32, 40]. | Moderate: significant variability in size and shape makes them less amenable to HTS than spheroids [57]. | Moderate: Some co-culture with immune cells is possible, but they often lack an autologous vascularized immune component [58]. | High: Patient-derived organoids (PDOs) allow for high-fidelity drug screening and capture the genetic landscape of the original tumor [9, 41]. | High human relevance, personalized medicine potential, recapitulates disease complexity [9, 32]. | High variability, standardization challenges, difficult integration with HTS, and a lack of a complete immune/stromal microenvironment [8, 57, 58]. |
| Organ-on-Chip (OoC)/microphysiological systems (MPS) | Very High: incorporates dynamic fluid flow, physiological mechanical forces, multi-cell co-cultures, and systemic effects [33, 60]. | Moderate to high: Throughput is improving with automation and standardized formats, but is not yet at traditional HTS levels [57]. | High: can simulate systemic effects and immune cell trafficking by connecting tissue compartments (e.g., tumor and lymph node) [51, 61]. | High: allows for dynamic TME emulation, multi-organ interactions (e.g., metabolism-toxicity), and advanced drug screening [33, 51]. | Mimics in vivo environment, uses human cells, allows real-time monitoring, enables multi-organ integration [33, 61]. | High cost, operational complexity, challenges in standardization and scale-up, complex data analysis [8]. |
During the preparation of this work, the authors used Napkin AI for creating the Figures and QuillBot for improving language clarity and grammar. No AI tools were used for scientific writing, data analysis, or interpretation. After using these tools, the authors thoroughly reviewed and edited all content and take full responsibility for the integrity and accuracy of the publication.
SC: Conceptualization, Writing—original draft, Visualization. AD: Methodology, Writing—original draft, Visualization. AS: Methodology, Writing—original draft. PZ: Writing—review & editing. NG: Writing—review & editing. RS: Writing—review & editing. VT: Writing—review & editing. LMA: Writing—review & editing, Supervision. DG: Conceptualization, Writing—review & editing, Supervision. All authors have read and approved the submitted version.
The authors declare no competing financial or non-financial interests related to the content of this manuscript.
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