Gene Network Based Differentiation of Stem Cells in a Microfabricated Array

基于基因网络的微加工阵列中干细胞的分化

• 批准号: 7872130
• 负责人: Yaakov Nahmias
• 金额: $ 5.4万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7872130
• 关键词: Advisory Committees; Animals; Arts; Award; Biomedical Engineering; Cell Count; Cell Line; Cell Lineage; Cells; Clinical; Collagen; Complex; Development; Development Plans; Developmental Biology; ES Cell Line; Elements; Endoderm; Endoderm Cell; Environment; Event; Gel; Gene Expression; General Hospitals; Genes; Genetic; Goals; Green Fluorescent Proteins; Growth Factor; Hepatic; Hepatocyte; Hormones; Human; In Vitro; Inflammatory Response; Institutes; K-Series Research Career Programs; Knowledge; Life; Liver; Liver Failure; Massachusetts; Mediator of activation protein; Microfabrication; Microfluidics; Molecular; Monitor; Mus; Network-based; Operative Surgical Procedures; Outcome; Pancreas; Pathway interactions; Phenotype; Population; Proliferating; Publishing; Regenerative Medicine; Reporter Genes; Research; Research Personnel; Resources; Signal Transduction; Source; Stem cells; Structure of beta Cell of islet; Techniques; Training; Work; career development; clinical application; cytokine; embryonic stem cell; insight; instructor; internal control; lectures; medical schools; novel; self-renewal; square foot; stem cell differentiation; tool; transcription factor

DESCRIPTION (provided by applicant): Dr. Yaakov Nahmias is a current Instructor in Surgery and Bioengineering at Harvard Medical School and seeks to establish himself as an independent investigator in the fields of regenerative medicine and BioMEMS. To that end, Dr. Nahmias submits this five years Career Development Award under the sponsorship of Dr. Mehmet Toner, director of the BioMEMS Resource Center and Dr. David Scadden, co- director of the Harvard Stem Cell Institute, both at Harvard Medical School. This award consists of a detailed career development plan which includes: 1) intensive training in microfabrication, 2) academic courses in Harvard and MIT, 3) seminars and lectures, and 4) advisory committee guidance. Research environment includes state-of-the-art facilities at Massachusetts General Hospital consisting of over 7000 sq ft of research and office space with adjacent cleanroom (class 1000), animal, and characterization facilities. The overall goal of this proposal is to elucidate and control the molecular mechanisms (transcription factor networks) underlying the differentiation of stem cells toward the hepatic phenotype. Our specific aims are to: 1) develop and characterize GFP-responsive mouse ES cell lines to major endodermal transcription factors; 2) microfabricate a living cell array for the culture of ES cells, and 3) elucidate and control the transcription factor networks regulating the differentiation of ES cell-derived endoderm toward the hepatic phenotype using soluble signals. We expect to: (a) develop multiple mouse embryonic stem cell lines with GFP- responsive transcription factor elements, (b) elucidate key genetic pathways underlying the differentiation of ES-derived endoderm toward the hepatic phenotype, and (c) provide new avenues to explore the differentiation potential of embryonic stem cells for scientific and clinical applications. One of the major challenges in the field today is the scarcity of functional human hepatocytes as the cells are notoriously difficult to proliferate in vitro. The treatment of liver failure would require around (10)10 hepatocytes. Treatments on this scale necessitate the development of alternative cellular sources that can potentially produce hepatocytes in high yields and large cell numbers.

描述（由申请人提供）： Yaakov Nahmias 博士现任哈佛医学院外科和生物工程讲师，致力于将自己打造成再生医学和 BioMEMS 领域的独立研究者。为此，Nahmias 博士在哈佛医学院 BioMEMS 资源中心主任 Mehmet Toner 博士和哈佛干细胞研究所联合主任 David Scadden 博士的赞助下提交了五年职业发展奖。该奖项由详细的职业发展计划组成，其中包括：1）微加工强化培训，2）哈佛大学和麻省理工学院的学术课程，3）研讨会和讲座，以及4）顾问委员会指导。研究环境包括马萨诸塞州总医院最先进的设施，包括超过 7000 平方英尺的研究和办公空间，以及相邻的洁净室（1000 级）、动物和表征设施。该提案的总体目标是阐明和控制干细胞向肝表型分化的分子机制（转录因子网络）。我们的具体目标是：1）开发并表征对主要内胚层转录因子有 GFP 反应的小鼠 ES 细胞系； 2) 微制造用于 ES 细胞培养的活细胞阵列，以及 3) 使用可溶性信号阐明和控制调节 ES 细胞衍生内胚层向肝表型分化的转录因子网络。我们期望：（a）开发多种具有 GFP 响应转录因子元件的小鼠胚胎干细胞系，（b）阐明 ES 衍生内胚层向肝表型分化的关键遗传途径，以及（c）提供新途径探索胚胎干细胞的分化潜力，用于科学和临床应用。当今该领域的主要挑战之一是功能性人类肝细胞的稀缺，因为这些细胞很难在体外增殖。肝衰竭的治疗需要大约 (10)10 个肝细胞。这种规模的治疗需要开发替代细胞来源，这些细胞来源有可能以高产量和大量细胞产生肝细胞。