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

3d打印（3DP）通过快速制造定制支架来增加组织工程（TE）的应用，其中精确的结构可以模拟天然组织。涉及多能干细胞、3d打印构建和细胞外基质（ECM）材料的TE应用已经变得越来越普遍。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。一项正在进行的fda和马里兰大学（UMD）合作开发并鉴定了一种细胞粘附离心（CAC）测定方法，以增强骨髓间充质干细胞（MSCs）的分离和分化。该建议将提高已建立的测定方法用于3D结构的实用性。它还将提供有用的洞察细胞-支架相互作用在三个维度，将通知标准化标准。这项工作将与马里兰大学和FDA的研究人员、教师和学生合作。适应3D支架设计的CAC检测的发展有可能通过提供一种机制来表征和纯化3D构建中的细胞，从而最大限度地减少TE治疗的翻译距离。细胞捕获装置将稳定3D支架，并允许基于整合素配体结合的非粘附细胞的表征。主要目标是：1)通过适应CAC实验，研究骨髓间充质干细胞的行为和从植入到3D构建体上的骨髓中纯化，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3D dp支架粘附和分化能力的多谱系分化潜力，研究MSCs与3D ECM聚合物支架的相互作用。3)研究使用CAC检测收集的非粘附细胞的捕获和表征。这将允许FDA进一步开发CAC检测，并评估其作为表征基于细胞成分3D构建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.