Directing embryonic stem cell (ESC)-derived hepatocytes is critical in understanding hepatic differentiation and applying cell-based treatment to severe liver diseases. While growth factor-based strategies are widely used, using chemical cues could present an alternative to optimize the strategies for stem cell differentiation. Here, for the first time, an inorganic calcium silicate (CS, CaSiO₃)-based approach, together with a modified four-stage differentiating protocol, was proposed to quantify the effects of CS extracts on inducing hepatic differentiation of human ESCs (H9 cells). The roles of CS-activated H9 cells in liver injury repair were tested by cell tracking and immunohistochemical staining. Results indicated that high concentrations of CS extracts initially enhance definitive endodermal (DE) lineage, followed by gradual DE differentiation at low CS concentrations. The order of CS addition is also crucial, since the presence at stemness stage and the absence at DE stage could optimize hepatic differentiation capacity of H9 cells, resulting in optimized cells that differentiate into functional hepatocyte-like cells. The addition of CS extracts at precursor hepatocyte stage enhances their maturity, which favors the turnover of liver injury in CCl₄-treated mice. These results provide an insight into applying bioactive inorganic biomaterials to foster hepatic differentiation of human ESCs for cell therapy.

指导胚胎干细胞（ESC）衍生的肝细胞对于理解肝脏分化并将基于细胞的治疗应用于严重的肝脏疾病至关重要。尽管基于生长因子的策略已被广泛使用，但使用化学提示可能是优化干细胞分化策略的替代方法。在这里，第一次是无机硅酸钙（CS，CaSiO ₃基于）的方法，以及修改后的四阶段分化方案，被提出来量化CS提取物对诱导人类ESC（H9细胞）肝分化的影响。通过细胞跟踪和免疫组织化学染色测试了CS活化的H9细胞在肝损伤修复中的作用。结果表明，高浓度的CS提取物最初会增强定型内胚层（DE）谱系，然后在低CS浓度下逐渐促进DE分化。CS的添加顺序也很关键，因为在干阶段存在和在DE阶段不存在可以优化H9细胞的肝分化能力，从而导致分化为功能性肝细胞样细胞的优化细胞。在前体肝细胞阶段添加CS提取物可增强其成熟度，₄处理的小鼠。这些结果为应用生物活性无机生物材料促进人类胚胎干细胞的肝分化以进行细胞治疗提供了见识。