An Enabling Technology for Human Cardiomyocyte Manufacturing

人类心肌细胞制造的使能技术

• 批准号: 10444010
• 负责人: Yuguo Lei
• 金额: $ 41.36万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10444010
• 关键词: 3-Dimensional; Address; Adopted; Alginates; Bioreactors; Blood Platelets; Caliber; Cardiac Myocytes; Cell Culture Techniques; Cell Death; Cell Line; Cell Therapy; Cells; Clinic; Clinical; Drug Screening; Erythrocytes; Funding; Generations; Goals; Growth; Growth Factor; Heart Diseases; Human; Hydrogels; In Vitro; Industry; Laboratory Research; Medicine; Methods; Minor; Mission; Modeling; Molecular Abnormality; National Heart, Lung, and Blood Institute; Persons; Phenotype; Production; Property; Protocols documentation; Reproducibility; Research; Safety; Small Business Innovation Research Grant; Small Business Technology Transfer Research; Source; Stress; Suspension Culture; Technology; Time; Tissue Engineering; Toxicity Tests; Tube; Variant; cell type; cost; cost effective; differentiation protocol; flasks; human disease; human pluripotent stem cell; in vivo; new technology; novel; operation; sound; stem cell differentiation

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.

项目摘要 人多能干细胞（human pluripotent stem cells，hPSC）及其衍生的心肌细胞（human pluripotent stem cells，hPSC-CMCs）是一种具有研究价值的细胞 和治疗人类疾病。然而，它们的广泛使用和进一步推进到诊所， 受限于大规模制造这些电池的困难。目前的细胞培养方法具有低的 效率低，不可扩展，并且只能以高成本小规模生产细胞。例如，使>10^10 每批hPSC-CMC仍然非常具有挑战性。此外，在产量和产量方面存在显著变化。 来自不同批次、实验室和公司的细胞的特性，导致不一致的研究和临床结果。 此外，产生的hPSC-CMC具有不成熟的表型，这限制了它们在体内和体外的使用。的 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。朝着这个 为了实现这一目标，该团队在初步研究中开发了一种可扩展的，可溶解的，细胞友好的3D微生物反应器。 问题研究微生物反应器为细胞提供了一个均匀的，可重复的，良好控制的，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， 在微生物反应器中扩增hPSC并使hPSC分化为心肌细胞。它的生存能力、成长能力 微生物反应器中细胞的速率、产率和质量提供了优于当前方法的巨大进步。体积 产量是目前最先进技术的250倍。新技术具有解决 大规模hPSC和hPSC-CMC制造挑战。拟议的研究将进一步发展和 积极验证技术和相关方法，使其稳健并为hPSC和hPSC做好准备- CMC制造。利用良好的初步研究和多样化的专家团队，具体目标是 （1）验证用于培养来自不同背景的hPSC的技术，以及（2）进一步开发和验证 高纯度hPSC-CMC的制备技术。完成这个项目，一个可扩展的，临床兼容的， 并且用于培养hPSC和hPSC-CMC的低成本细胞培养技术将可用。该技术可以 也可用于培养与National Heart相关的其他细胞（如血小板和来自hPSC的红细胞）， 肺和血液研究所的使命。这项技术正在使并将大大推进心脏医学 疾病