HiDef B8: Commercialization and scaled production of defined, robust, and cost-effective media for iPSCs

研究及相关其他项目信息 Defined Bioscience，Inc. 7.项目摘要 用于细胞培养的生长培养基的不一致性是工业和学术界的一个重要问题， 几十年来一直困扰着实验室。在生物制药领域，基于细胞和细胞衍生的疗法正以惊人的速度增长。 这是一个巨大的速度，需要明确的，动物的零部件，以满足FDA和cGMP法规。是 因此，对于用于细胞培养的无血清培养基的需求和采用的增加并不奇怪 增长归因于其卓越的批次间一致性和降低的不必要的风险 动物/异生物质污染物。这对人类诱导多能干细胞（hiPSC）， 快速发展的技术，广泛的研究和临床应用。这些细胞呈现出许多 与以前的技术相比的优势，包括人类起源，形成多类细胞的能力， 系统和组织，相对容易的生产和修改，以及功能相关性。坚固耐用- 当培养用于下游应用的hiPSC时，所表征的方案是极其重要的。高 多能性、遗传稳定性、大规模生产和维持功能都必须牢记， hiPSC。然而，尽管现在可用于iPSC的一系列确定成分的培养基，但这些培养基中的许多未能满足要求。 hiPSC研究的全部需求，包括多次传代的稳健性和实验需求，低成本， 易于制作，周末无需更换介质，组件定义齐全。这些特征对于健壮， 在hiPSC领域进行一致和负担得起的研究。 就在今年，我们的Defined Bioscience团队在一个大型的实验室中对常见的成分确定培养基成分进行了经验性测试。 一系列浓度和组合，分离出一个定义明确的配方，具有高稳健性， > 100次细胞传代，并在无周末传代计划中保持成功。该媒体被称为B8， 科学顾问委员会成员保罗·伯里奇的实验室达到或超过了第一阶段SBIR的预期 发展建议。在这里，我们将扩展这项工作，以准备高清晰度B8，制定B8最终确定为最佳 组分浓度和降低的成本。这种优化的配方将在学术界和 美国的工业实验室，以确认简化和应用的hiPSC培养的稳健性和有效性 和方法。我们打算使HiDef B8成为一种负担得起的、确定的培养基，用于在全球范围内培养强大的hiPSC。 高传代次数和无周末背景下，显著降低hiPSC的成本和复杂性 跨领域的技术。这将使稳定的细胞培养和良好表征的起始点成为可能。 随后的差异化和应用hiPSC技术的努力，同时降低的障碍， 进入现场进行hiPSC开发。

RESEARCH & RELATED Other Project Information Defined Bioscience, Inc. 7. PROJECT SUMMARY Inconsistency of growth media for cell culture is a significant problem in both industry and academia that has plagued laboratories for decades. In biopharma, cell-based and cell-derived therapies are growing at a tremendous rate and require well-defined, animal-free components to meet FDA and cGMP regulations. It is therefore not surprising that there is an increased need for and adoption of serum-free media for cell culture growth that is attributed to its superior batch-to-batch consistency and reduced risk of unwanted animal/xenobiotic contaminants. This is particularly critical to human induced pluripotent stem cells (hiPSCs), a rapidly evolving technology with wide research and clinical application. These cells present numerous advantages compared to previous technologies, including human origin, the ability to form multi-class cell systems and tissues, relative ease of production and modification, and functional relevance. Robust and well- characterized protocols are of utmost importance when culturing hiPSCs for downstream applications. High pluripotency, genetic stability, large-scale production and maintained function all must be kept in mind for hiPSCs. Yet despite the array of defined media now available for iPSCs, many of these media fail to satisfy the full needs of hiPSC research, including robustness over multiple passages and experimental needs, low cost, ease of production, weekend-free media changes and fully defined components. Such traits are critical for robust, consistent and affordable research in the hiPSC space. Just this year, our Defined Bioscience team empirically tested common defined media components over a large array of concentrations and combinations, isolating a well-defined recipe with high robustness, efficacy over >100 cell passages, and maintained success in a weekend-free passaging schedule. This media, termed B8 by the lab of scientific advisory board member Paul Burridge, met or exceeded the expectations for a Phase I SBIR development proposal. Here we will extend this work to prepare HiDef B8, a formulation of B8 finalized to optimal component concentrations and reduced cost. This optimized formulation will be tested across academic and industrial labs in the United States to confirm robustness and efficacy in streamlined and applied hiPSC culture and methodologies. We intend to make HiDef B8 an affordable, defined media for robust hiPSC culture across high passage numbers and in a weekend-free context, significantly reducing the cost and complexity of hiPSC technologies across the field. This will enable stable cell culture and a well-characterized starting point for subsequent differentiation and applied hiPSC technology efforts, while simultaneously lowering the barrier of entry for hiPSC development in the field.