Development of hepatocytes from ES cells

ES细胞发育成肝细胞

• 批准号: 7264009
• 负责人: VALERIE B GOUON-EVANS
• 金额: $ 12.95万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7264009
• 关键词: Acetoacetates; Activities of Daily Living; Address; Blood Vessels; Cell Differentiation process; Cell Separation; Cells; Characteristics; Condition; Data; Development; Ectoderm; Embryo; Endoderm; Exposure to; Generations; Germ Layers; Goals; Hematopoietic; Hepatic; Hepatocyte; Human; Hydrolase; Insulin; Intestines; Kinetics; Liver; Liver Failure; Lung; Mediating; Mesoderm; Modeling; Mus; Muscle; Neurons; Organ; Pancreas; Population; Pre-Clinical Model; Regulation; Replacement Therapy; Research Personnel; Serum; Staging; System; Thyroid Gland; Tissues; Translating; Translations; Transplantation; activin A; cell type; embryonic stem cell; human embryonic stem cell; mouse model; progenitor; programs; research study; stem

DESCRIPTION (provided by applicant): The development of definitive endoderm and its subsequent specification leads to the formation of many of the major organs including the liver, pancreas, lungs, thyroid, and intestines. Under appropriate culture conditions, mouse embryonic stem (ES) cells differentiate into embryoid bodies (EBs) that contain derivatives of all three germ layers. Most of these studies have focused on the development of mesoderm and ectoderm derivatives. The development of hematopoietic, vascular, muscle and neuronal lineages were found to parallel that in the early embryo, thus validating the ES system for early developmental studies. More recently, studies have provided evidence for endoderm development in ES differentiation cultures and demonstrated the generation of insulin expressing cells and cells with hepatocyte characteristics. ES studies have not yet established conditions for the efficient induction of endoderm or its specification to liver. This proposal addresses these issues by investigating specific steps of development including (1) induction of ES cells to an endoderm fate, (2) specification of endoderm into a hepatocyte progenitor, called the hepatoblast and (3) maturation of the hepatoblast into a functional hepatocyte. Thus, the liver is the focus in this application as an endoderm-derived tissue. Early endoderm fated cells as well as the hepatoblast will be characterized from ES cell cultures using cell sorting analysis. The liver repopulation potential of endoderm-derived cells will be analyzed in two mouse models of liver failure. The project will also seek to translate the murine ES cell differentiation program to optimize the generation of human hepatocytes. Therefore, the ultimate goal of these studies is to develop a feasible approach to generate functional hepatocytes from ES ells and to understand the mechanisms that orchestrate the hepatocyte development from ES cells.

描述（由申请人提供）： 定形内胚层的发育及其随后的规格导致许多主要器官的形成，包括肝脏、胰腺、肺、甲状腺和肠。在适当的培养条件下，小鼠胚胎干（ES）细胞分化成含有所有三个胚层衍生物的胚状体（EB）。这些研究大部分集中在中胚层和外胚层衍生物的开发上。造血、血管、肌肉和神经谱系的发育被发现与早期胚胎中的发育平行，从而验证了 ES 系统用于早期发育研究。最近，研究为 ES 分化培养中的内胚层发育提供了证据，并证明了胰岛素表达细胞和具有肝细胞特征的细胞的产生。 ES研究尚未建立有效诱导内胚层或其向肝脏分化的条件。 该提案通过研究发育的具体步骤来解决这些问题，包括 (1) 诱导 ES 细胞走向内胚层命运，(2) 将内胚层规范化为肝细胞祖细胞，称为肝母细胞，以及 (3) 肝母细胞成熟为功能性肝细胞。因此，肝脏作为内胚层来源的组织是本申请的焦点。将使用细胞分选分析从 ES 细胞培养物中表征早期内胚层命运细胞以及肝母细胞。将在两种肝衰竭小鼠模型中分析内胚层衍生细胞的肝脏再生潜力。该项目还将寻求转化小鼠 ES 细胞分化程序，以优化人类肝细胞的生成。因此，这些研究的最终目标是开发一种可行的方法来从 ES 细胞生成功能性肝细胞，并了解协调 ES 细胞发育为肝细胞的机制。