An Enabling Technology for Human Cardiomyocyte Manufacturing

人类心肌细胞制造的使能技术

• 批准号: 10626105
• 负责人: Yuguo Lei
• 金额: $ 39.59万
• 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/10626105
• 关键词: 3-Dimensional; Acceleration; Address; Adopted; Alginates; Bioreactors; Blood Platelets; Cardiac Myocytes; Cell Culture Techniques; Cell Death; Cell Line; Cell Therapy; Cells; Clinic; Clinical; Cultured Cells; Diameter; Drug Screening; Erythrocytes; Friends; Funding; Generations; Goals; Growth; Growth Factor; Heart Diseases; Human; Human body; 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; Tissue Engineering; Toxicity Tests; Tube; Variant; cell type; cost; cost effective; differentiation protocol; flasks; human disease; human pluripotent stem cell; improved; in vivo; manufacture; new technology; novel; operation; sound; stem cell expansion; technology validation

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.

项目概要 人类多能干细胞 (hPSC) 及其衍生心肌细胞 (hPSC-CMC) 对于研究具有重要价值 然而，目前它们的广泛使用和进一步推进到临床。 受限于大规模生产这些细胞的难度，目前的细胞培养方法水平较低。 效率低，不可扩展，并且只能以高成本生产小规模的电池，例如制造>10^10。 此外，每批次的 hPSC-CMC 仍然非常具有挑战性。 不同批次、实验室和公司的细胞特性不同，导致研究和临床结果不一致。 此外，产生的 hPSC-CMC 具有不成熟的表型，限制了它们在体内和体外的使用。 该项目的总体目标是开发一种新型细胞培养技术来应对这一挑战。 为了实现这一目标，该团队在前期开发了一种可扩展、可溶解且对细胞友好的3D微生物反应器 微生物反应器为细胞提供了均匀、可重复、良好控制和友好的环境。 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 并将 hPSC 分化为心肌细胞 活力、生长。 微生物反应器中细胞的速率、产量和质量比现有方法取得了巨大进步。 产量是当前最先进技术的 250 倍，新技术具有解决该问题所需的所有功能。 大规模 hPSC 和 hPSC-CMC 制造挑战将进一步发展。 大力验证该技术和相关方法，使其稳健并为 hPSC 和 hPSC 做好准备 利用良好的初步研究和多元化的专家团队，具体目标是 (1) 验证培养不同背景 hPSC 的技术，以及 (2) 进一步开发和验证 制备高纯度 hPSC-CMC 的技术完成了该项目，这是一种可扩展的、临床兼容的、 用于培养 hPSC 和 hPSC-CMC 的低成本细胞培养技术将可用。 也可用于培养与国家心脏相关的其他细胞（例如来自 hPSC 的血小板和红细胞）， 肺和血液研究所的使命是推动并显着推动心脏医学的发展。 疾病。