An Enabling Technology for Human Cardiomyocyte Manufacturing

项目总结 人多能干细胞(HPSCs)及其来源的心肌细胞(hPSC-CMCs)具有重要的研究价值 以及治疗人类疾病。然而，它们的广泛使用和进一步发展到临床目前 受到大规模制造这些电池的困难的限制。目前的细胞培养方法较低。 效率低，不可扩展，只能生产小规模的电池，成本高。例如，制作&gt；10^10 每批HPSC-CMCs仍然具有很大的挑战性。此外，产量和产量也存在显著差异。 不同批次、实验室和公司的细胞特性，导致研究和临床结果不一致。 此外，生产的hPSC-CMCs具有不成熟的表型，这限制了它们在体内和体外的使用。这个 该项目的总体目标是开发一种新的细胞培养技术来应对这一挑战。朝向这个方向 为了实现这一目标，该团队在初步开发出了一种可扩展、可溶解、对细胞友好的3D微生物反应器 学习。微生物反应器为细胞提供了统一、可重复、可控和友好的 微环境，导致极高的培养效率和一致性。该团队已经证明了 在微生物反应器中扩增hPSCs并将其分化为心肌细胞。生存能力、增长能力 微生物反应器中细胞的速率、产量和质量都比目前的方法有了很大的进步。体积学 收益率是目前最先进水平的250倍。这项新技术具有所有必要的功能，以解决 大规模hPSC和hPSC-CMCS制造的挑战。拟议的研究将进一步发展和 大力验证技术和相关方法，使其强大并为hPSC和hPSC做好准备- CMCS制造。利用完善的初步研究和不同的专家团队，具体目标是 以(1)验证不同背景的hPSCs培养技术和(2)进一步开发和验证 高纯度hPSC-CMCs的制备技术。完成这个项目，一个可扩展的，临床兼容的， 并将提供培养hPSC和hPSC-CMCs的低成本细胞培养技术。这项技术可以 也可用于培养与国家心脏相关的其他细胞(如来自hPSCs的血小板和红细胞)， 以及血液研究所的使命。这项技术是可行的，并将显著促进心脏药物的发展 疾病。

PROJECT SUMMARY Human pluripotent stem cells (hPSCs) and their derived cardiomyocytes (hPSC-CMCs) are valuable for studying and treating human diseases. However, their widespread use and further advancement to clinics are currently limited by the difficulty of manufacturing these cells at large scales. The current cell culture methods have low efficiency, are not scalable, and can only produce cells at small scales with high cost. For instance, making>10^10 hPSC-CMCs per batch is still very challenging. Additionally, there are significant variations in the yield and properties of cells from different batches, labs, and companies, leading to inconsistent research and clinic results. Further, the produced hPSC-CMCs have immature phenotypes that limit their use both in vivo and in vitro. The overall goal of this project is to develop a novel cell culture technology to address this challenge. Toward this goal, the team has developed a scalable, dissolvable, and cell-friendly 3D microbioreactor in the preliminary studies. The microbioreactor provides cells a uniform, reproducible, well-controlled, and friendly microenvironment, resulting in extremely high culture efficiency and consistency. The team has demonstrated expanding hPSCs and differentiating hPSCs into cardiomyocytes in the microbioreactor. The viability, growth rate, yield, and quality of cells in the microbioreactor offer large advances over current methods. The volumetric yield is 250 times of current state-of-the-art. The new technology has all the needed features to address the large-scale hPSCs and hPSC-CMCs manufacturing challenge. The proposed research will further develop and vigorously validate the technology and the associated methods to make it robust and ready for hPSCs and hPSC- CMCs manufacturing. Leveraging sound preliminary studies and a diverse team of experts, the specific aims are to (1) validate the technology for culturing hPSCs from diverse backgrounds and (2) further develop and validate the technology for preparing high-purity hPSC-CMCs. Completing this project, a scalable, clinically compatible, and low-cost cell culture technology for culturing hPSCs and hPSC-CMCs will be available. The technology can also be used for culturing other cells (such as platelets and red blood cells from hPSCs) related to National Heart, Lung, and Blood Institute’s mission. The technology is enabling and will significantly advance medicine for heart disease.