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

1265029 -米勒从培养的巨核细胞中产生3000 - 6000亿单位的血小板将需要在造血（血液）干细胞和祖细胞扩增、巨核细胞产生和成熟以及血小板释放和恢复方面取得进展。鼻窦血流的剪切对血小板前体和血小板释放是重要的。受骨髓窦壁龛的启发，该项目将采用一个微生物反应器，其两个隔间由多孔支架隔开，多孔支架涂有带有粘附配体的微孔聚乙二醇水凝胶。为了模拟通过窦内皮形成血小板前体，将巨核细胞播种在上腔室的水凝胶上，并通过下腔室灌注培养基。微生物反应器将用于研究剪切应力、水凝胶刚度、黏附配体类型和密度对血小板前体和血小板生成的影响。将研究增加pH梯度和趋化因子以进一步增加血小板形成。灌注系统将允许在血小板释放时收集血小板，这很重要，因为血小板在20-24°C储存期间质量会恶化，在37°C储存时甚至更快。培养源血小板的质量将通过激动剂诱导激活来验证。微生物反应器可以用于临床生产血小板，使用中空纤维生物反应器和螺旋缠绕膜盒等格式来产生所需的表面积。在美国和欧洲，每年有数百万单位的血小板被输注。利用来自患者或匹配的血液干细胞的巨核细胞在培养中生产血小板将增加供应，降低污染风险和不良免疫反应，但目前在培养中生产血小板的效率非常低。这项研究的成功完成将使培养源血小板的大规模生产更接近于实现。从事该项目的研究生和本科生将在跨学科的环境中接受培训。该项目的一个关键要素是让研究生为高中生开发有关微流体、聚合物水凝胶、巨核细胞和血小板的教育模块，包括为周六癌症学院提供的两节半天的课程和在当地高中的演讲。该奖项来自CBET部门的生物技术、生化和生物工程项目，由材料研究部门的生物材料项目共同主办。

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.