Embryonic Stem Cell Research Career Enhancement

胚胎干细胞研究职业提升

• 批准号: 6806938
• 负责人: Kenneth S. Zuckerman
• 金额: $ 23.07万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6806938
• 关键词: biological signal transduction; bone morphogenetic proteins; cell biology; cell differentiation; human embryonic stem cell line; pluripotent stem cells; protein protein interaction; protein structure function

DESCRIPTION (provided by applicant): The purpose of this proposal is for Dr. Zuckerman to obtain intensive training in embryonic stem (ES) cell biology. He will train in the laboratory of Dr. Martin Pera in the Institute for Reproduction and Development at Monash University in Melbourne, Australia. Dr. Pera is a pioneer in human ES cell technology and a contributer of nearly half of the human ES cell lines that are approved by NIH and are currently available. All of the training and experiments done under this grant will utilize these NIH-approved ES cell lines (ES01- ES06). Training will include technical training in isolation, identification, and maintenance of ES cells in culture, techniques for inducing ES cell differentiation into cells of desired lineages, and methods of extrinsic DNA transfer into ES cells. There will be extensive guided and self-driven reading of ES cell and related scientific and ethical literature. There will be frequent discussions with Dr. Pera, weekly lab meetings for discussion of recent results, and frequent seminars on topics relevant to ES cell biology. There is a rich stem cell research environment in Melbourne. Hematopoietic stem cell biology pioneer Don Metcalf and his extended group are at the Walter and Eliza Hall Institute, and Paul Simmons and his group at the Peter MacCallum Cancer Institute work on stem cell plasticity. Until recently, human ES cells could only be grown and maintained in the undifferentiated pluripotent state over a mouse embryo fibroblast feeder cell layer. Bone Morphogenic Protein (BMP)-2 is produced by the ES cells and/or early differentiated progeny, especially under conditions of stress. BMP-2 drives differentiation of ES cells into endodermal cells. Gremlin, a DAN family member, which is upregulated by BMP-2, has been shown recently to be a relatively specific antagonist of BMP-2 and BMP-4. In preliminary studies, Dr. Pera found that feeder cells in ES cell cultures produce gremlin transcripts. We will test the hypothesis that gremlin, by interfering with BMP-2 function, may play a large role in the ability of the feeder cells to maintain ES cells in the undifferentiated state. We will examine the ability of gremlin to prevent ES cell differentiation in cultures with no feeder cells and the ability of "impotent" feeder cells to support ES cell self-renewal and growth after being transfected with gremlin cDNA and overexpressing gremlin protein. We also will determine whether inhibition of gremlin production abolishes the ability of a functional feeder cell layer to maintain ES cells in the undifferentiated state.

描述（由申请人提供）： 该提案的目的是让Zuckerman博士获得胚胎茎（ES）细胞生物学的强化培训。他将在澳大利亚墨尔本莫纳什大学的繁殖与发展研究所的马丁·佩拉博士的实验室培训。 Pera博士是人类ES细胞技术的先驱，也是由NIH批准并目前可用的近一半人类ES细胞系的贡献者。根据该赠款进行的所有培训和实验将利用这些NIH批准的ES细胞系（ES01- ES06）。培训将包括隔离，鉴定和培养中ES细胞维持的技术训练，诱导ES细胞分化为所需谱系细胞的技术以及外在DNA转移到ES细胞中的方法。 ES细胞以及相关的科学和道德文献将有广泛的指导和自我驱动阅读。将与Pera博士，每周实验室会议进行讨论，以讨论最近的结果，并经常讨论与ES细胞生物学相关的主题。墨尔本有丰富的干细胞研究环境。造血干细胞生物学先驱Don Metcalf及其扩展小组在Walter and Eliza Hall Institute，Paul Simmons及其小组在Peter MacCallum Cancer Cancer Institute工作，从事干细胞可塑性。直到最近，人类ES细胞只能在小鼠胚胎成纤维细胞喂食器细胞层上生长和维持在未分化的多能状态。骨形态发生蛋白（BMP）-2由ES细胞和/或早期分化后代产生，尤其是在压力条件下。 BMP-2将ES细胞分化为内皮细胞。 DAN家庭成员Gremlin被BMP-2上调，最近已被证明是BMP-2和BMP-4的相对特定的拮抗剂。在初步研究中，Pera博士发现ES细胞培养物中的饲养细胞会产生绿色转录本。我们将检验以下假设：通过干扰BMP-2功能，Gremlin可能在馈线细胞保持在未分化状态下的ES细胞的能力中起着重要作用。我们将检查灰色的能力，以防止没有进料细胞的培养物中的ES细胞分化以及“无能”进料细胞在被gremlin cDNA转染和过表达的gremlin蛋白后支持ES细胞自我更新和生长的能力。我们还将确定抑制灰色生产是否消除了功能性馈线细胞层保持ES细胞在未分化状态下的能力。