中文摘要：

背景：在胚胎干细胞向肝细胞诱导分化的培养条件基础上增加向胆管上皮细胞分化的培养条件，使其向胆管上皮细胞方向分化在理论上是可行的。 目的：验证在细胞生长因子胚胎干细胞体外定向分化为胆管上皮细胞的可行性。设计：单一样本观察。 单位：中山大学附属第一医院。 材料：选用BALB／c系小鼠胚胎干细胞（BALB／c—ES）由中山大学实验动物中心建系和保存。分化生长因子、酸性成纤维细胞生长因子、肝细胞生长因子、上皮生长因子、角化细胞生长因子均为美国Sigma公司产品，实验用抗小鼠细胞角蛋白7（CK7）和细胞角蛋白19（CK19）购自丹麦DAKO公司，间接免疫荧光试剂盒为美国Biodesign产品，IX70-S8F2倒置相差荧光显微镜为日本OLYMPUS产品。 方法：实验于2004—10／2005-06在中山大学附属第一医院中心实验室完成。①将小鼠胚胎干细胞进行拟胚体分化，在培养系统中按不同时间段分别添加分化生长因子、酸性成纤维细胞生长因子、肝细胞生长因子、上皮生长因子、角化细胞生长因子，使胚胎干细胞向胆管上皮细胞方向分化。以培养液中不添加上述细胞生长因子为对照组，使胚胎干细胞自然分化。②采用倒置相差荧光显微镜动态观察细胞生长及三维的环状结构形成情况。③于细胞分化第10天开始采用免疫细胞化学方法检测胆管上皮细胞标记物CK7、CK19，γ-谷胺酰转肽酶（GGT）表达变化，同时观察γ-谷胺酰转肽酶阳性细胞的形态学特点。 主要观察指标：①细胞生长及三维环状结构形成情况。②胆管上皮细胞标记蛋白CK7、CK19的表达。③胆管上皮细胞标记酶GGT表达及其阳性细胞形态学情况。结果：①细胞生长及三维的环状结构形成情况：胚胎干细胞悬浮培养10h后即可见小的拟胚体形成并悬浮在培养液中。分化第10天在拟胚体细胞分?

英文摘要：

BACKGROUND： Previous research demonstrated that embryonic stem （ES） cells can be induced to differentiate into hepatocytes. It is feasible to induce ES cells to differentiate into biliary epithelial （BE） cells by specific induction conditions based on the previous research. OBJECTIVE： To validate the feasibility to induce ES cells to differentiate into BE cells in the presence of cell growth factors. DESIGN： Single sample observation. SETTING： First Hospital of Sun Yat-sen University. MATERIALS： Undifferentiated BALB/C-ES cell line was provided by the Experimental Animal Center of Sun Yat-sen University; Transforming growth factor, acidic fibroblast growth factor, hepatocyte growth factor, epidermal growth factor, and keratinocyte growth factor by Sigma, USA. First antibody： anti-mouse cytokeratin 7 （CK7） and CK19 by DAKO, Denmark; indirect immunofluorescence from Biodesign, USA; and inverted contrast fluorescence microscope by OLYMPUS IX70-S8F2, Japan. METHODS： The experiment was performed at Central Laboratory of the First Affiliated Hospital of Sun Yat-sen University from October 2004 to June 2005. During the culture of embryonic bodies （EBs） derived from ES cells, some growth factors such as Transforming growth factor, acidic fibroblast growth factor, hepatocyte growth factor and epidermal growth factor were respectively added into medium to induce BE cells differentiation. ES cells in culture condition with no added growth factors served as control. The differentiation status and three-dimensional duct-like structure formation of ES cells was observed dynamically by inverted microscope. The markers of BE cells such as CK7, CK19 and gamma-glutamyltransferase （GGT） were detected by immunocytochemistry and GGT-positive cells were observed. MAIN OUTCOME MEASURES： （2）Cell growth and three-dimensional duct-like structure formation; （2）expressions of the marker proteins of BE cells such as CK7 and CKIg;（3）expression of GGT, the marker enzyme of BE cells, and