Establishment of a cryo-bank of lineage-committed neural progenitor cells produced from engineered human pluripotent stem cells

建立由工程化人类多能干细胞产生的谱系定向神经祖细胞冷冻库

• 批准号: NC/X002144/1
• 负责人: Tilo Kunath
• 金额: $ 25.79万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=NC%2FX002144%2F1
• 关键词: Establishment; cryo; bank; lineage; committed

Human induced pluripotent stem cells (iPSCs), reported in 2007 by Shinya Yamanaka, can be derived from blood or skin of any individual. They are unique because they can be grown indefinitely in the lab, and they have the capacity to produce any mature cell type - a property known as pluripotency. These attributes make iPSCs ideal candidates to replace animals in biological research. The rationale is that iPSC-derived heart cells, nerve cells, liver cells and other cell types can be used for many experiments instead of mice or other animals. Although the complexity of iPSC cultures is not yet approaching that of a whole animal, there is significant progress towards replacing animals in many areas of research. Despite iPSCs being discovered over 10 years ago, there have been barriers to their wide-spread adoption in academia and industry, which have prevented them from achieving their potential as an alternative to animal use in research. The first barrier was acquisition of high-quality iPSC lines - this is now routine and 1000s of iPSC lines are available world-wide. The second barrier facing many labs is reliable and consistent production of mature specialised cell types (such as cardiac or nerve cells) from iPSCs. The inherent pluripotent nature of iPSCs means they are primed to produce all cell types in a dish. The protocols developed to corral and coax iPSCs to produce specific desired cell types, such as neurons, are complicated, lengthy and sensitive to minor perturbations. Furthermore, published "differentiation" protocols that work for particular iPSC lines are often not easily transferable to other iPSC lines. This has led to a significant level of frustration in the field, and has restricted iPSC technology to selected labs with the necessary expertise and resources to overcome the challenges of iPSC differentiation. The main aim of this proposal is to completely remove this barrier and increase confidence in iPSC technology through the establishment of the Edinburgh Progenitor Cell Bank (EPB). We have established methods to differentiate iPSCs along cell lineage pathways and freeze them while in a progenitor cell state. A "progenitor cell" is a transition cell type that is half-way between an iPSC and a mature cell type. The frozen progenitor cells we produce are restricted and committed to produce a specific mature cell type after thawing, and they can be cultured by any laboratory and does not require any iPSC expertise. The EPB will supply and support the use of lineage-committed neural progenitor cells for research purposes in academia and industry. Each batch of frozen progenitor cells undergo test-thaws and vigorous quality-control checks to ensure that recipient labs are supplied with high-quality functional cells. The EPB will be for the neuroscience community in the first instance, but can be expanded to other areas of biological research when other frozen iPSC-derived progenitor cells (eg. cardiac, liver) are included. The goal of the EPB is to provide diverse labs, particularly heavy animal users, with ready-to-use specialised human cells as a reliable and effective replacement for animal research subjects. Supplying progenitor cells in this way increases the capacity for use of this animal alternative by removing the need for labs to possess the skills and knowledge necessary to control stem cell differentiation.

Shinya Yamanaka 于 2007 年报道，人类诱导多能干细胞 (iPSC) 可以源自任何个体的血液或皮肤。它们是独一无二的，因为它们可以在实验室中无限期地生长，并且有能力产生任何成熟的细胞类型——这种特性被称为多能性。这些属性使 iPSC 成为生物研究中替代动物的理想候选者。其基本原理是，iPSC 衍生的心脏细胞、神经细胞、肝细胞和其他细胞类型可以代替小鼠或其他动物用于许多实验。尽管 iPSC 培养物的复杂性尚未接近整个动物的复杂性，但在许多研究领域在替代动物方面已经取得了重大进展。尽管 iPSC 已在 10 多年前被发现，但其在学术界和工业界的广泛采用仍存在障碍，这阻碍了它们在研究中发挥其作为动物用途替代品的潜力。第一个障碍是获取高质量的 iPSC 细胞系 - 这现在已成为惯例，全球有数千条 iPSC 细胞系。许多实验室面临的第二个障碍是从 iPSC 可靠且一致地生产成熟的特殊细胞类型（例如心脏或神经细胞）。 iPSC 固有的多能性质意味着它们能够在培养皿中产生所有细胞类型。为圈养和诱导 iPSC 产生特定所需细胞类型（例如神经元）而开发的方案复杂、冗长且对微小扰动敏感。此外，已发布的适用于特定 iPSC 系的“分化”方案通常不容易转移到其他 iPSC 系。这导致了该领域的巨大挫败感，并且将 iPSC 技术限制在具有必要专业知识和资源的选定实验室，以克服 iPSC 差异化的挑战。该提案的主要目的是通过建立爱丁堡祖细胞库（EPB）来彻底消除这一障碍并增强对 iPSC 技术的信心。我们已经建立了沿细胞谱系途径分化 iPSC 并在祖细胞状态下冷冻它们的方法。 “祖细胞”是介于iPSC和成熟细胞类型之间的过渡细胞类型。我们生产的冷冻祖细胞受到限制，解冻后致力于产生特定的成熟细胞类型，并且它们可以由任何实验室培养，不需要任何 iPSC 专业知识。 EPB 将提供并支持谱系定向神经祖细胞用于学术界和工业界的研究目的。每批冷冻祖细胞都经过试解冻和严格的质量控制检查，以确保为受体实验室提供高质量的功能细胞。 EPB 首先将面向神经科学界，但当包括其他冷冻 iPSC 衍生的祖细胞（例如心脏、肝脏）时，可以扩展到生物研究的其他领域。 EPB 的目标是为不同的实验室（特别是重型动物用户）提供现成的专用人类细胞，作为动物研究对象的可靠且有效的替代品。以这种方式提供祖细胞，通过消除实验室需要掌握控制干细胞分化所需的技能和知识，增加了使用这种动物替代品的能力。