Comparison of the proposed AM-Based 3D Invasion Models vs. Conventional Assays.
| Feature | Proposed AM-Based 3D Model | Matrigel Invasion Assay [31] | Transwell Assay (2D/Pseudo-3D) [32] | In Vivo Tail Vein Injection Model [33] |
|---|---|---|---|---|
| ECM Source | Human-derived amniotic membrane (AM) | Murine tumor ECM (Matrigel) | Synthetic filters + ECM coating | Native mouse tissues (no matrix scaffold used) |
| Structural Integrity | Preserved basement membrane and stroma | Gel-like, lacks defined structure | Rigid, artificial support | Physiological ECM of lung/liver |
| Biochemical Composition | Collagens I/IV, laminin, fibronectin + growth factors | Variable protein concentration of murine origin (9–12 mg/mL) + tumor-derived factors | Essentially absent unless coated (collagen/fibronectin added in µg/cm2) | Full physiological composition, but not human-derived |
| Mechanical Properties | Elastic modulus: 0.1–10 kPa; tensile strength 1–5 MPa | Elastic modulus: 100–500 Pa; lacks tensile integrity | Artificially rigid; tensile strength > 10 MPa; stiffness far above physiological range | Elastic modulus varies widely (e.g., lung ~1 kPa, liver ~0.5–1.5 kPa, bone > 10 MPa) |
| Batch Consistency | Moderate; donor screening required | High batch variability | High | High consistency per mouse strain |
| Real-Time Imaging Compatibility | High (live-cell imaging, confocal, multiphoton) | Limited due to opacity | High (endpoint only) | Limited; requires intravital microscopy or bioluminescence |
| EMT and Invasion Modeling | Supports EMT in 3D with native ECM cues | EMT-inducing, but less structured | Limited EMT modeling | Full EMT and metastasis cascade, but not trackable early |
| Co-culture Capability | Supports stromal, immune, endothelial cells | Limited by gel stability | Low | Possible but complex (e.g., bone marrow transplant) |
| Patient-Derived Organoid Use | Feasible | Difficult | Poor integration | Not directly applicable |
| Translational Relevance | High; human-derived ECM and structure | Moderate; murine origin | Low | High biological control, but low experimental control |
| Cost and Availability | Low-cost; clinically sourced | High cost; proprietary | Inexpensive | High; requires animal facility |
| Ethical & Regulatory Barrier | Minimal (clinical waste) | Moderate | None | High (animal use protocols required) |
| Throughput | Medium (scalable with inserts or chips) | Low | High | Low |
| Quantitative Readouts | Invasion depth, EMT markers, protease activity | Invasion area/number | Cell count or fluorescence | Tumor burden (bioluminescence, histology) |