Streamlined Banking of Human Pluripotent Stem Cells Through Molecular Authenticat

通过分子验证简化人类多能干细胞的存储

• 批准号: 7802771
• 负责人: JONATHAN M. AUERBACH
• 金额: $ 22.78万
• 依托单位: GLOBALSTEM, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7802771
• 关键词: Address; Adult; Attitude; Autologous; Biological Assay; CD34 gene; Cell Culture Techniques; Cell Line; Cell Therapy; Cells; Collaborations; Communities; Complex; Development; Floods; Future; G-Banding; Gene Expression; Hematopoietic; Human; Immunophenotyping; In Vitro; Institution; Karyotype; Karyotype determination procedure; Letters; Maintenance; Mesenchymal Stem Cells; Molecular; Mutation; Pharmacotherapy; Phase; Phenotype; Play; Pluripotent Stem Cells; Process; Property; Quality Control; Research; Role; Running; Scheme; Scientist; Seeds; Sickle Cell; Source; Standardization; Stem Cell Research; Stem cells; System; Testing; Time; Translational Research; Umbilical Cord Blood; Undifferentiated; Universities; Vision; Work; base; cell bank; cost effective; human disease; human embryonic stem cell; in vitro Model; induced pluripotent stem cell; interest; pluripotency; public health relevance; stability testing

DESCRIPTION (provided by applicant): The recent creation of human induced pluripotent stem cells (iPSC) from reprogrammed or de-differentiated adult cells has changed the approach and attitude of many to pluripotent stem cell research. These new iPSC are potentially a valuable source of in vitro models of complex, polygenic human diseases. They may find an important role in the discovery of new drugs and therapies. They are also a potential downstream source of cells for autologous cell therapy. However, these new cells are still subject to some of the same roadblocks that have been standing in the way of advancing human embryonic stem cell (hESC) research. These cells are inherently difficult to maintain in culture, they require meticulous quality control and standardization, and they must be made readily available to the research community. Cell culture has played a central and critical role in the advancement of both basic and translational research. As a result, scientists and institutions have come to recognize the advantages of maintaining cell banks in order to support the development and maintenance of healthy cell lines. In this application, we propose to create a streamlined human pluripotent stem cell (hPSC - hESC and iPSC) banking system in which we will optimize the authentication and quality control (QC) processes of hPSC lines. Human iPSC and human ESC share many common properties - including culture requirements, marker expression, and pluripotency. We propose to use three human iPSC lines as proof-of-principle for hPSC banking. These are potentially interesting and important cell lines, especially for the hematopoietic and mesenchymal stem cell fields. This system will be cost effective and will strive for self-sufficiency. We will greatly enhance the turnaround time for making the banks of cells available by employing a characterization scheme mainly comprised of molecular assays. The proposed assays are less expensive (requiring less cellular material) and quicker (having shorter run times) than traditional assays. We will validate the new assays by comparing the results with those obtained by traditional tests being used now - including immunophenotyping and G-band karyotyping. Once accomplished, we will have taken a large step forward toward realizing the full potential of these stem cells. This is a forward-looking application with a vision to the near future need for banking hundreds of hPSC lines. PUBLIC HEALTH RELEVANCE: In this proposal we are laying the groundwork for an hPSC bank that will be able to respond quickly to the impending flood of hESC and iPSC lines. By validating a set of molecular authentication assays we will have taken the first step toward an efficient banking system. The ultimate endpoint is an hPSC bank that can quickly and economically accession and make important cell lines available to the research community.

描述（由申请人提供）：最近从重编程或去分化的成体细胞中创建的人类诱导多能干细胞（iPSC）改变了许多人对多能干细胞研究的方法和态度。这些新的iPSC是复杂的多基因人类疾病体外模型的潜在有价值的来源。他们可能会在发现新药物和新疗法方面发挥重要作用。它们也是用于自体细胞治疗的细胞的潜在下游来源。然而，这些新细胞仍然受到一些阻碍人类胚胎干细胞（hESC）研究的障碍。这些细胞本质上很难在培养中维持，它们需要细致的质量控制和标准化，并且必须随时提供给研究界。细胞培养在基础研究和转化研究的发展中起着核心和关键的作用。因此，科学家和机构已经认识到维持细胞库的优势，以支持健康细胞系的开发和维护。在本申请中，我们提出创建流线型的人多能干细胞（hPSC - hESC和iPSC）库系统，其中我们将优化hPSC系的鉴定和质量控制（QC）过程。人iPSC和人ESC具有许多共同的特性-包括培养要求、标记物表达和多能性。我们建议使用三种人iPSC细胞系作为hPSC库的原理证明。这些是潜在的有趣和重要的细胞系，特别是造血和间充质干细胞领域。这一系统将具有成本效益，并将努力实现自给自足。我们将通过采用主要由分子测定组成的表征方案，大大提高细胞库的周转时间。所提出的测定比传统测定更便宜（需要更少的细胞材料）和更快（具有更短的运行时间）。我们将通过与现在使用的传统检测方法（包括免疫表型分析和G带核型分析）的结果进行比较来验证新的检测方法。一旦完成，我们将朝着实现这些干细胞的全部潜力迈出一大步。这是一个前瞻性的应用程序，其愿景是在不久的将来需要银行数百个hPSC行。 公共卫生相关性：在这项提案中，我们正在为hPSC银行奠定基础，该银行将能够迅速应对即将到来的hESC和iPSC生产线的洪水。通过验证一套分子鉴定试验，我们将朝着高效的银行系统迈出第一步。最终的终点是一个hPSC库，它可以快速、经济地加入并向研究界提供重要的细胞系。