单克隆来源的小鼠胎肝HPP-CFC肝上皮分化潜能的研究  

Differentiation Potential of Clonal HPP-CFC Into Hepatic Lineage in Developing Mouse Fetal Liver

作　　者：周军年[1] 王韫芳[1] 要晖宇[2] 贺文艳[2] 陈海旭[1] 李思霆[1] 史岩[1] 施双双[1] 南雪[1] 白慈贤[1] 刘兵[2] 岳文[1] 毛宁[3] 裴雪涛[1] 

机构地区：[1]军事医学科学院输血医学研究所干细胞与再生医学研究室,北京100850 [2]军事医学科学院附属307医院肿瘤学研究室,北京100071 [3]军事医学科学院基础医学研究所细胞生物学研究室,北京100850

出　　处：《生物化学与生物物理进展》2009年第7期830-839,共10页Progress In Biochemistry and Biophysics

基　　金：国家高技术研究发展计划(863)(2006AA02A107);国家重点基础研究发展规划(973)(2005CB522702);国家自然科学基金重点项目(30730043);国家自然科学基金(30671098)资助项目~~

摘　　要：为研究胎肝中造血和肝上皮发育的关系,建立了小鼠胎肝高增殖潜能集落形成细胞(HPP-CFC)培养体系,并进行了单克隆培养以及诱导分化实验.在造血和肝诱导因子的共同作用下,对单克隆来源的HPP集落细胞向造血和肝上皮细胞进行诱导分化,采用透射电镜(TEM)、巢式RT-PCR、细胞免疫荧光检测,从细胞形态、超微结构、上皮细胞分化标志等方面对分化后的细胞进行检测.检测结果显示,诱导后的部分细胞具有肝细胞特异性的超微结构,并不同程度地表达白蛋白(ALB)、甲胎蛋白(AFP)、细胞角蛋白(CK8,CK18)等肝上皮分化标志,同时还表达间质标志α-SMA和血管内皮细胞标志Flk-1.免疫磁珠分选表明:胎肝来源的HPP-CFC主要来自于CD45+细胞,CD45-细胞不具有形成造血细胞克隆的能力.在肝上皮细胞分化潜能上,流式分选获得的CD49f+/Sca-1+细胞与未分选细胞无明显差异.该模型的克隆源性通过细胞混合实验进行证明.研究结果表明,改进的胎肝来源的HPP-CFC可能代表了一个新的造血细胞向肝上皮细胞分化的单克隆模型,为研究胎肝中造血和非造血细胞的发育关系提供了一个新的切入点.To investigate the hepatic development association with hematopoiesis, a high proliferative potential colony forming cells （HPP-CFC） model of mouse fetal liver was set up. Some differentiational assays based on individual HPP colonies were performed. Under the condition of combinations of hematopoietic and hepatic factors, some individual HPP colonies were induced into hematopoietic and hepatic cells, which were examined with transmission electron microscope （TEM）, nested RT-PCR and immunofluorescence staining. The results showed that induced HPP colonies cells with a specific ultrastructure similar to hepatic epithelial cells, expressed hepatic markers including albumin （ALB）, α-fetoprotein （AFP）, cytokeratins （CK8, CK18） at different extent of percentage. These cells also expressed mesenchymal marker α-SMA and primary endothelial cell marker Flk-1. The MACS results suggested that the fetal liver-derived HPP-CFCs are all from CD45^＋ cells, while CD45^- cells have no capacity to form hematopoietic colony at all. The FACS sorted CD49f^＋/Sca-1^＋ ceils have no difference of hepatic differentiation potential compared with whole fetal liver cells. The clonality was confirmed by cell mixing assay. Taken together, the HPP-CFC may represent a novel clonal model for hepatic differentiation from the blood cells in the mouse feta liver and will shed light on the associations underlying the hepatic and hematopoietic development.

关 键 词：胎肝 高增殖潜能集落形成细胞 肝上皮细胞 造血 发育 

分 类 号：Q2[生物学—细胞生物学] Q132.4