RHESUS ES CELLS: A MODEL TO STUDY PANCREAS DEVELOPMENT

恒河猴 ES 细胞：研究胰腺发育的模型

• 批准号: 6381948
• 负责人: Jon S Odorico
• 金额: $ 14.4万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-15 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6381948
• 关键词: Macaca mulatta; animal tissue; biological models; cell differentiation; embryonic stem cell; gene expression; histogenesis; in situ hybridization; laboratory mouse; model design /development; northern blottings; pancreas; polymerase chain reaction

A comprehensive knowledge of the genetic programs and embryonic inductive events regulating human pancreas development would advance our understanding of congenital pancreatic malformations and aid in developing rational treatment strategies for diabetes mellitus. Significant insights to the mechanisms of pancreatic organogenesis have been gained through recent experiments in mice and chickens. However, critical differences in early developmental pathways between these species and humans places the direct applicability of these data to human development in question. Furthermore, functional studies in human development are restricted by the ethical implications of directly manipulating human embryos. Consequently, a more "human" model based on nonhuman primates would have broad application for studying human development. Recently, embryonic stem (ES) cell lines from nonhuman primates (rhesus macaque and common marmoset) have been derived and their potential for differentiation into derivatives of all three embryonic germ layers (ectoderm, mesoderm, and endoderm) in vivo has been characterized. Their capacity for lineage-restricted differentiation in vitro, however, has not been thoroughly tested to date. in contrast, neural, hematopoietic and muscle lineage differentiation of mouse ES cells in vitro has been extensively studied. Recently, the ability of mouse ES cells to undergo elements of endoderm and pancreatic differentiation in culture has also been demonstrated. Therefore, we will test the hypothesis that pancreatic endocrine and/or exocrine lineage specification can be recapitulated in rhesus ES cells induced to differentiate. The specific aims of this proposal are: l) To determine the expression pattern of endoderm related genes as well as pancreatic exocrine/endocrine-specific genes in rhesus ES cells differentiating in vitro, and 2) To determine the potential of rhesus ES cells for pancreatic lineage differentiation in ES cell derived tumors in immunocompromised mice. A combination of RT-PCR analysis, Northern hybridization, ribonuclease protection assay, in situ hybridization, and immunofluorescence studies will be used to characterize the pattern of pancreatic gene expression. These studies will provide a basis for the refinement of an in vitro primate model of human pancreatic development. Such a model system would have important implications for investigating the mechanisms controlling pancreatic differentiation and for developing new cell-based strategies for treating diabetes.

对调节人类胰腺发展的遗传计划和胚胎归纳事件的全面知识将提高我们对先天性胰腺畸形的理解，并有助于制定糖尿病的合理治疗策略。通过在小鼠和鸡的最新实验中获得了对胰腺器官发生机制的重要见解。但是，这些物种与人之间的早期发育途径的关键差异使这些数据直接适用于相关的人类发展。此外，人类发展中的功能研究受到直接操纵人类胚胎的伦理意义的限制。因此，基于非人类灵长类动物的更“人类”模型将在研究人类发展方面具有广泛的应用。最近，已经得出了来自非人类灵长类动物的胚胎（ES）细胞系（ES）细胞系（恒河猕猴和公共果果），并将其分化为体内所有三个胚胎胚层（Ectoderm，中胚层和内胚层）的衍生物的潜力已经表征。然而，迄今为止，尚未对其在体外谱系限制分化的能力。相反，已经广泛研究了小鼠ES细胞的神经，造血和肌肉谱系的分化。最近，也已经证明了小鼠ES细胞在培养中经历内胚层和胰腺分化元件的能力。因此，我们将检验以下假设：胰腺内分泌和/或外分泌谱系可以在诱发分化的恒河生细胞中概括。该提案的具体目的是：l）确定内胚层相关基因的表达模式以及在体外区分的恒龙ES细胞中的胰腺外分泌/内分泌特异性基因，以及2），以确定恒河类ES细胞对胰腺的潜力。在免疫功能低下的小鼠中，ES细胞衍生肿瘤的谱系分化。 RT-PCR分析，北部杂交，核糖核酸酶保护测定，原位杂交和免疫荧光研究的结合将用于表征胰腺基因表达的模式。这些研究将为人类胰腺发育的体外灵长类动物模型提供改进的基础。这种模型系统将对控制胰腺分化的机制和制定新的基于细胞的糖尿病策略具有重要意义。