NSF/FDA Scholar In Residence: 3D Cell Adhesion Assay for Cellularized Scaffold Characterization and Enhancement

摘要：Fisher，John提案编号：3D打印（3DP）通过快速制造定制支架来增加组织工程（TE）应用，其中精密架构可以模拟天然组织。涉及多能干细胞、3DP构建体和细胞外基质（ECM）材料的TE应用变得越来越普遍。为了将这些技术转化为治疗方法，有必要了解3D环境中的细胞行为。正在进行的FDA-马里兰州大学（UMD）合作已经导致细胞粘附离心（CAC）测定的开发和表征，以增强从骨髓中分离和分化间充质干细胞（MSC）。该提议将增强已建立的用于3D构建体的测定的实用性。它还将提供有用的洞察细胞支架相互作用的三维，将告知标准化标准。 这项工作将与马里兰州大学和FDA的研究人员、教职员工和学生合作。CAC检测的开发可适应3D支架设计，通过提供一种表征和纯化3D结构内细胞的机制，有可能最大限度地减少TE治疗的平移距离。细胞捕获装置将稳定3D支架，并允许基于整合素-配体结合来表征非粘附细胞。主要目标是：1）通过CAC测定的调整研究MSC行为和从接种到3D构建体上的骨髓中的纯化，其中将评估离心力、孵育时间和3D构建体结构的影响：2）通过检查ECM添加剂对3DP支架粘附和分化能力的多谱系分化潜力来研究MSC与3D ECM-聚合物支架的相互作用：和3）研究使用CAC测定收集的非粘附细胞的捕获和表征。这将使FDA能够进一步开发CAC检测，并评估其作为表征基于细胞组分3D构建体的疗法的方法的实用性。

ABSTRACTPI: Fisher, JohnProposal Number: 16410873D Printing (3DP) is increasing tissue engineering (TE) applications through rapid manufacture of custom scaffolds wherein precision architectures can mimic native tissue. TE applications involving multipotent stem cells, 3DP constructs, and extracellular matrix (ECM) materials have become increasingly prevalent. In order to translate these technologies as therapeutics it is necessary to understand cell behavior within the 3D environment. An ongoing FDA-University of Maryland (UMD) collaboration has led to the development and characterization of a cell adhesion centrifugation (CAC) assay to enhance isolation and differentiation of mesenchymal stem cells (MSCs) from bone marrow. This proposal will enhance the utility of the established assay for use with 3D constructs. It will also provide useful insight into cell-scaffold interaction in three dimensions that will inform standardization criteria. The work will partner researchers, faculty and students at the University of Maryland and the FDA.The development of the CAC assay to accommodate 3D scaffold designs has the potential to minimize the translational distance for TE therapies by providing a mechanism to characterize and purify cells within 3D constructs. The cellular capture device will stabilize 3D scaffolds and allow characterization of non-adherent cells based on integrin-ligand binding. The key aims are: 1) investigate MSC behavior and purification from bone marrow seeded onto 3D constructs through adaptation of the CAC assay wherein the impact of centrifugation force, incubation time, and 3D construct architecture will be evaluated: 2) investigate the interaction of MSCs with 3D ECM-polymer scaffolds through examination of multi-lineage differentiation potential enabled by ECM additives to 3DP scaffold adhesion and differentiation capacity: and 3)investigate the capture and characterization of non-adhered cells collected using the CAC assays. This will allow the FDA to further develop the CAC assay and to evaluate its utility as a method to characterize the cellular component 3D construct based therapies.