The extracellular matrix (ECM) in a liver-specific extracellular matrix (L-ECM) scaffold facilitates hepatocyte viability and maintains hepatocyte functions in vitro. However, whether an intact composition of ECM is required for an efficient ECM-based substrate design remains to be clarified. In this study, two L-ECM hydrogels, namely L-ECM I and L-ECM II, were prepared by pepsin solubilization at 4 °C and 25 °C, respectively. The solubility at 4 °C was 50% whereas that at 25 °C was 95%, thus indicating well-preserved L-ECM. Analysis confirmed higher ECM protein components (especially collagen) in L-ECM II, along with denser fiber network and larger fiber diameter. L-ECM II gel exhibited high compression strength and suitable viscoelastic properties. Furthermore, hepatocytes in L-ECM II showed higher expression of liver-specific functions in 3D culture and wider spread while maintaining the cell-cell contacts in 2D culture. Therefore, an intact L-ECM is important to realize effective substrates for liver tissue engineering.

肝脏特异性细胞外基质（L-ECM）支架中的细胞外基质（ECM）可促进肝细胞活力并在体外维持肝细胞功能。然而，高效的基于 ECM 的基板设计是否需要完整的 ECM 成分仍有待澄清。在本研究中，分别在4℃和25℃下通过胃蛋白酶溶解制备了两种L-ECM水凝胶，即L-ECM I和L-ECM II。4°C 时的溶解度为 50%，而 25°C 时的溶解度为 95%，因此表明 L-ECM 保存良好。分析证实，L-ECM II 中的 ECM 蛋白质成分（尤其是胶原蛋白）更高，纤维网络更致密，纤维直径更大。L-ECM II 凝胶表现出高压缩强度和合适的粘弹性能。此外，L-ECM II 中的肝细胞在 3D 培养中表现出更高的肝脏特异性功能表达，并且在 2D 培养中保持细胞间接触的同时分布更广泛。因此，完整的L-ECM对于实现肝组织工程的有效底物非常重要。