Two-compartment microfluidic bioreactor with functionalized PEG hydrogels to promote platelet production in culture

具有功能化 PEG 水凝胶的两室微流体生物反应器可促进培养中血小板的产生

• 批准号: 1265029
• 负责人: William Miller
• 金额: $ 45万
• 依托单位: Northwestern University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-01 至 2017-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1265029&HistoricalAwards=false
• 关键词: Two; compartment; microfluidic; bioreactor; functionalized

1265029 - MillerGenerating many units of 300-600 billion platelets from megakaryocytic cells in culture will require advances in hematopoietic (blood) stem and progenitor cell expansion, megakaryocyte production and maturation, and platelet release and recovery. Shear from blood flow in the sinuses is important for proplatelet and platelet release. Inspired by the bone marrow sinusoid niche, this project will employ a microbioreactor with two compartments separated by a porous support coated with microporous poly(ethylene glycol) hydrogel functionalized with adhesive ligands. To mimic proplatelet formation across the sinusoidal endothelium, megakaryocytes will be seeded on the hydrogel in the upper compartment and medium will be perfused through the lower compartment. The microbioreactor will be used to investigate effects of shear stress, hydrogel stiffness, and adhesive ligand types and density on proplatelet and platelet production. Increasing gradients of pH and chemokines will be investigated to further increase proplatelet formation. The perfusion system will allow platelets to be harvested as they are released, which is important because platelet quality deteriorates during storage at 20-24°C and even faster at 37°C. The quality of culture-derived platelets will be verified via agonist-induced activation. The microbioreactor could be scaled for clinical production of platelets using formats such as hollow-fiber bioreactors and spiral-wound membrane cartridges to generate the required surface area. Several million units of platelets are transfused each year in the United States and Europe. Production of platelets in culture by megakaryocytes derived from patient or matched blood stem cells would increase the supply and decrease the contamination risk and adverse immune responses, but platelet production in culture is currently very inefficient. Successful completion of the proposed research will bring the large-scale production of culture-derived platelets closer to fruition. Graduate students and undergraduates working on this project will be trained in an interdisciplinary environment. A key element of the project is for the graduate students to develop educational modules on microfluidics, polymer hydrogels, megakaryocytic cells, and platelets for high school students, including two half-day classes for the Oncofertility Saturday Academy and presentation at local high schools.The award from the Biotechnology, Biochemical, and Bioengineering Program of the CBET Division is co-sponsored by the Biomaterials Program of the Division of Materials Research.

1265029-MILLER从培养中的巨核细胞生成3,000-6,000亿个单位的血小板将需要在造血(血液)干细胞和祖细胞扩增、巨核细胞的生产和成熟以及血小板的释放和恢复方面取得进展。鼻窦血流的剪切力对血小板原和血小板的释放很重要。受骨髓正弦波生态位的启发，该项目将使用一个具有两个隔室的微生物反应器，由一个涂有微孔聚乙二醇水凝胶的多孔支撑体分隔，微孔聚乙二醇水凝胶由粘合剂配体功能化。为了模拟跨越血窦内皮细胞的原血小板形成，巨核细胞将被种植在上部隔室的水凝胶上，而培养液将通过下部隔室灌流。微生物反应器将被用来研究剪切力、水凝胶硬度、黏附配体的类型和密度对原血小板和血小板产生的影响。增加pH和趋化因子的梯度将被研究以进一步增加前血小板的形成。灌流系统将允许在血小板释放时获取它们，这一点很重要，因为在20-24°C的存储过程中，血小板质量会恶化，在37°C时会更快。培养衍生的血小板的质量将通过激动剂诱导的激活来验证。利用中空纤维生物反应器和螺旋缠绕膜盒等形式，可以将微生物反应器扩大到临床生产血小板，以产生所需的表面积。在美国和欧洲，每年有数百万个单位的血小板被输注。从患者或配对的血液干细胞来源的巨核细胞在培养中产生血小板将增加供应，降低污染风险和不良免疫反应，但目前在培养中产生血小板的效率非常低。拟议研究的成功完成将使培养衍生血小板的大规模生产更接近成果。从事这个项目的研究生和本科生将在跨学科的环境中接受培训。该项目的一个主要内容是让研究生为高中生开发关于微流体、聚合物水凝胶、巨核细胞和血小板的教育模块，包括肿瘤生育星期六学院的两节半天课程和在当地高中的演讲。CBET部门生物技术、生化和生物工程项目的奖项由材料研究部生物材料项目共同赞助。