Summary
This application note demonstrates how ECM stiffness regulates hepatic iron biology in C3A liver spheroids. Compared with 2D culture and Matrigel®, LunaX™ ECM supported more uniform spheroids and stiffness-dependent increases in TFR1, TFR2 and MT2 expression across 0.5–4 kPa conditions.
What you’ll learn
- How to establish C3A hepatic spheroids across 0.5, 2 and 4 kPa LunaX ECM conditions.
- How this stiffness range models the mechanical transition from healthy liver parenchyma to early fibrotic tissue.
- How ECM stiffness affects hepatic spheroid formation, regularity and uniformity over six days.
- How TFR1, TFR2 and MT2 expression changes between 2D monolayers, Matrigel and tunable 3D ECM.
- How a mechanically defined liver model can support research into iron overload, haemochromatosis, MASLD and liver fibrosis.
Key findings
- C3A cells formed well-defined spherical aggregates in LunaX, while 2D cultures remained flat and Matrigel produced loose, irregular aggregates.
- Spheroid regularity and uniformity increased with LunaX ECM stiffness, with the most consistent morphology observed at 4 kPa.
- TFR1 expression increased progressively across 0.5–4 kPa LunaX conditions and was highest at 4 kPa.
- TFR2 and MT2 were substantially upregulated in LunaX compared with 2D and Matrigel, with the strongest responses at higher stiffness.
- The coordinated stiffness-dependent response of TFR1, TFR2 and MT2 was absent or markedly attenuated in conventional 2D and non-tunable Matrigel conditions.
