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获得功能性的肝细胞可用于研究肝细胞再生机理,并可为细胞移植和药物筛选提供生物材料。该研究采用四步法诱导多能干细胞分化成肝细胞,比较传代和不传代分化细胞的形态和肝细胞特异标志的差异表达。结果发现,利用诱导多能干细胞分化可获得白蛋白(albumin,ALB)阳性的成熟肝细胞,不传代法获得的细胞更接近于肝细胞形态,两组均获得甲胎蛋白(alphafetoprotein,AFP)和ALB阳性表达的细胞;GATA6(GATA binding protein 6)和HNF4A(hepatocyte nuclear factor 4 alpha)表达水平没有显著性差异,但不传代法分化细胞的AFP和ALB表达量高于传代法,存在显著性差异。结果提示,细胞因子与细胞间、多细胞间相互作用等因素影响多能干细胞的分化,细胞间相互作用有利于肝细胞定向诱导分化过程中肝细胞的成熟。
Functional hepatocytes can be used to study the mechanism of hepatocyte regeneration and provide biomaterials for cell transplantation and drug screening. In this study, four-step method was used to induce pluripotent stem cells to differentiate into hepatocytes. The morphological differences between differentiated cells and non-differentiated cells and hepatocyte-specific markers were compared. The results showed that albumin (ALB) -positive mature hepatocytes could be obtained by inducing pluripotent stem cell differentiation. The cells obtained by the non-passage method were more close to the hepatocyte morphology. Both groups obtained alphafetoprotein (AFP) and ALB positive cells; there was no significant difference between GATA6 and HNF4A (P <0.05), but the expression levels of AFP and ALB in differentiated cells without passaging were higher than that in passaging, there was a significant difference . The results suggest that factors such as cytokines and intercellular and multicellular interactions affect the differentiation of pluripotent stem cells, and cell-cell interactions are conducive to the maturation of hepatocytes during the differentiation-induced differentiation of hepatocytes.