人外周血单个核细胞向肝样细胞转化的体外诱导培养方法

背景：外周血单个核细胞在体外条件下可向肝样细胞转化。目的：探索体外诱导外周血单个核细胞向肝样细胞分化的培养方法。方法：采用密度梯度离心法联合贴壁法纯化肝硬化患者外周血单个核细胞,以含巨噬细胞集落刺激因子、白细胞介素3及β-巯基乙醇培养基培养6d后,诱导组用含肝细胞生长因子,成纤维细胞生长因子4的培养基培养14d;以未诱导培养的单个核细胞及L02肝脏细胞系为对照。结果与结论：外周血单个核细胞呈透明圆形、类圆形,用巨噬细胞集落刺激因子、白细胞介素3及β-巯基乙醇培养基培养6d后,部分细胞向成纤维样细胞生长,诱导后部分细胞呈多边形,多角形,14d后细胞形态逐渐接近肝细胞,并表达白蛋白、甲胎蛋白、角蛋白18。说明在一定的诱导条件下,外周血单个核细胞在体外可以向肝样细胞分化。

In vitro induction method for the differentiation of human peripheral blood mononuclear cells into hepatocyte-like cells

BACKGROUND：Peripheral blood mononuclear cells can differentiate into hepatocyte-like cells in vitro.OBJECTIVE：To explore the induced method that peripheral blood mononuclear cells can be differentiated to hepatocyte-like cells in vitro.METHODS：Peripheral blood mononuclear cells were isolated from patients by using density gradient centrifugation and cells adherent method,and cultured in the culture medium containing recombinant human macrophage colony stimulating factor（M-CSF）,interleukin 3（IL-3） and β-Mercaptoethanol for 6 days.The purified cells were cultured with hepatocyte growth factor and fibroblast growth factor 4 to accelerate induction for 14 days（experimental group）.Non-induced mononuclear cells and L02 liver cell line were used as controls.RESULTS AND CONCLUSIONS：Freshly isolated peripheral blood mononuclear cells were small round,uniform size,strong refraction.After induction with M-CSF,IL-3 and β-Mercaptoethanol for 6 days,the part of cells grew into fibroblasts;and after induction with HGF and FGF-4 for 14 days,the cell morphology was similar to liver cells,and the cells expressed albumin,alpha-fetoprotein,cytokeratin 18.Peripheral blood mononuclear cells can be transformed into liver cells in vitro under certain induced conditions.