Characterization of a cell line for use as a reference standard for human embryon

用作人类胚胎参考标准的细胞系的表征

• 批准号: 7676596
• 负责人: JONATHAN M. AUERBACH
• 金额: $ 6.64万
• 依托单位: GLOBALSTEM, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-15 至 2009-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7676596
• 关键词: Applications Grants; Cell Line; Commercial Sources; Communities; Condition; Data; Development; Human; Human Cloning; Human Development; Laboratories; Lead; Personal Satisfaction; Property; Qualifying; Quality Control; Reference Standards; Research Personnel; Side; Stem Cell Research; Undifferentiated; Work; base; cell bank; human embryonal carcinoma cell; human embryonic stem cell; human embryonic stem cell line; novel therapeutics; research study; scale up

DESCRIPTION (provided by applicant): Human embryonic stem cell (hESC) research has a great potential to further our understanding of basic mechanisms of human development and may lead to the development of novel therapeutics. Absent from this young field has been the acceptance and use of reference standard cell lines. Uniform use of a reference standard would permit investigators to reliably compare results across laboratories and would aid in building sequential advances in the field. A reference cell line should be easy to obtain; the cell line should be relatively easy to grow and maintain; there should be available data for the comparison of results; data and experiments should be easily reproducible; and comparable results using the cell line should be obtained across experiments and across laboratories. In addition to these properties, a reference standard should represent, as closely as possible, the normal or experimental cell line and a particular state of that cell line. In this grant application, we are proposing a reference standard for the undifferentiated hESC state. We propose that nullipotent 2102Ep is better representative of the undifferentiated hESC state than any non-hESC line. Currently, however, there is no commercial source of standardized 2102Ep. We propose to: 1. Perform side-by-side characterization and comparison of two clones of the human embryonal carcinoma cell line 2102Ep 2102Ep cl.4D3 and 2102Ep cl.2A6 - in order to determine which clone is best suited as a reference standard for the undifferentiated hESC state. The criteria used to determine which clone is best will be based on elevated, uniform expression of markers of the undifferentiated state. 2. Moderately scale-up best clone as determined in Specific Aim 1 to a working cell bank; characterize working cell bank and compare results to those obtained in Aim 1. Characterization will examine the identity, stability, and marker expression of the two clones. The significance of this work is its general applicability to hESC research and the benefit that will be reaped by the hESC research community once such a qualified reference standard is widely available.The end result and relevance of this project is a clone of human EC 2102Ep which closely represents the undifferentiated human ESC state. This clone will be produced under standardized conditions; it will be well-characterized, easy to use, and readily available. Availability of this clone is a necessary step to hasten the pace of sequential advances in hESC research.

描述（由申请人提供）：人类胚胎干细胞（hESC）研究具有极大的潜力，可以进一步加深我们对人类发育基本机制的理解，并可能导致新疗法的开发。缺乏从这个年轻的领域一直是接受和使用的参考标准细胞系。统一使用参考标准将使研究人员能够可靠地比较各实验室的结果，并有助于在该领域取得连续的进展。参考细胞系应易于获得;细胞系应相对易于生长和维持;应具有可用于结果比较的数据;数据和实验应易于重现;使用细胞系进行的实验和实验室之间的结果应具有可比性。除这些特性外，参比标准品应尽可能接近正常或实验细胞系以及该细胞系的特定状态。在这个资助申请中，我们提出了一个未分化hESC状态的参考标准。我们认为，nullipotent 2102Ep比任何非hESC系更能代表未分化的hESC状态。然而，目前还没有标准化的2102Ep的商业来源。我们建议：1.对人胚胎癌细胞系2102 Ep、2102 Ep cl.4D3和2102 Ep cl.2A6的两个克隆进行并行表征和比较，以确定哪个克隆最适合作为未分化hESC状态的参考标准。用于确定哪个克隆是最佳的标准将基于未分化状态的标志物的升高的、均匀的表达。2.将特定目标1中确定的最佳克隆适度放大至工作细胞库;表征工作细胞库并将结果与目标1中获得的结果进行比较。表征将检查两个克隆的同一性、稳定性和标志物表达。这项工作的意义在于它对hESC研究的普遍适用性，以及一旦这样一个合格的参考标准被广泛使用，hESC研究团体将获得的好处。该克隆将在标准化条件下生产;它将具有良好的特性，易于使用，并且随时可用。这种克隆的可用性是加快hESC研究连续进展步伐的必要步骤。