Abstract：
Hepatic microenvironment plays an essential role in liver regeneration, providing the necessary conditions for cell proliferation, differentiation and tissue rearrangement. One of the key factors for hepatic tissue reconstruction is the extracellular matrix (ECM), which through collagenous and non-collagenous proteins provide a three-dimensional structure that confers support for cell adhesion and assists on their survival and maintenance. In this scenario, placental ECM may be eligible for hepatic tissue reconstruction, once these scaffolds hold the major components required for cell support. Therefore, this preliminary study aimed to access the possibility of mouse embryonic stem cells differentiation into hepatocyte-like cells on placental scaffolds in a three-dimensional dynamic system using a Rotary Cell Culture System. Following a four-phase differentiation protocol that simulates liver embryonic development events, the preliminary results showed that a significant quantity of cells adhered and interacted with the scaffold through outer and inner surfaces. Positive immunolabelling for alpha fetus protein and CK7 suggest presence of hepatoblast phenotype cells, and CK18 and Albumin positive immunolabelling suggest the presence of hepatocyte-like phenotype cells, demonstrating the presence of a heterogeneous population into the recellularized scaffolds. Periodic Acid Schiff-Diastase staining confirmed the presence of glycogen storage, indicating that differentiate cells acquired a hepatic-like phenotype. In conclusion, these preliminary results suggested that mouse placental scaffolds might be used as a biological platform for stem cells differentiation into hepatic-like cells and their establishment, which may be a promissing biomaterial for hepatic tissue reconstruction.
摘要：
肝微环境在肝再生中起着至关重要的作用，为细胞增殖提供必要条件，分化和组织重排。肝组织重建的关键因素之一是细胞外基质（ECM），它通过胶原和非胶原蛋白提供三维结构，为细胞粘附提供支持，并有助于它们的存活和维持。在这种情况下，胎盘ECM可能符合肝组织重建的条件，一旦这些支架保持细胞支持所需的主要组件。因此,这项初步研究旨在使用旋转细胞培养系统在三维动态系统中在胎盘支架上获得小鼠胚胎干细胞分化为肝细胞样细胞的可能性。遵循模拟肝脏胚胎发育事件的四期分化方案，初步结果表明，大量细胞通过外表面和内表面与支架粘附并相互作用。α胎儿蛋白和CK7的阳性免疫标记提示存在肝细胞表型细胞，CK18和白蛋白阳性免疫标记提示肝细胞样表型细胞的存在，证明在再细胞化的支架中存在异质群体。周期性酸性Schiff-Distase染色证实了糖原储存的存在，表明分化细胞获得肝样表型。总之,这些初步结果表明，小鼠胎盘支架可能作为干细胞分化为肝样细胞的生物平台及其建立，这可能是一种用于肝组织重建的生物材料。
