Di Buduo Christian A, Wray Lindsay S, Tozzi Lorenzo, Malara Alessandro, Chen Ying, Ghezzi Chiara E, Smoot Daniel, Sfara Carla, Antonelli Antonella, Spedden Elise, Bruni Giovanna, Staii Cristian, De Marco Luigi, Magnani Mauro, Kaplan David L, Balduini Alessandra
Department of Molecular Medicine, University of Pavia, Pavia, Italy; Biotechnology Research Laboratories, Istituto di Ricovero e Cura a Carattere Scientifico San Matteo Foundation, Pavia, Italy;
Department of Molecular Medicine, University of Pavia, Pavia, Italy; Biotechnology Research Laboratories, Istituto di Ricovero e Cura a Carattere Scientifico San Matteo Foundation, Pavia, Italy; Department of Biomedical Engineering, Tufts University, Medford, MA;
Blood. 2015 Apr 2;125(14):2254-64. doi: 10.1182/blood-2014-08-595561. Epub 2015 Jan 9.
We present a programmable bioengineered 3-dimensional silk-based bone marrow niche tissue system that successfully mimics the physiology of human bone marrow environment allowing us to manufacture functional human platelets ex vivo. Using stem/progenitor cells, megakaryocyte function and platelet generation were recorded in response to variations in extracellular matrix components, surface topography, stiffness, coculture with endothelial cells, and shear forces. Millions of human platelets were produced and showed to be functional based on multiple activation tests. Using adult hematopoietic progenitor cells our system demonstrated the ability to reproduce key steps of thrombopoiesis, including alterations observed in diseased states. A critical feature of the system is the use of natural silk protein biomaterial allowing us to leverage its biocompatibility, nonthrombogenic features, programmable mechanical properties, and surface binding of cytokines, extracellular matrix components, and endothelial-derived proteins. This in turn offers new opportunities for the study of blood component production ex vivo and provides a superior tissue system for the study of pathologic mechanisms of human platelet production.
我们展示了一种可编程的生物工程三维丝基骨髓龛组织系统，该系统成功模拟了人类骨髓环境的生理学，使我们能够在体外制造功能性人类血小板。利用干/祖细胞，记录了巨核细胞功能和血小板生成对细胞外基质成分、表面形貌、硬度、与内皮细胞共培养以及剪切力变化的反应。通过多次激活测试表明，该系统产生了数百万功能性人类血小板。利用成人造血祖细胞，我们的系统展示了再现血小板生成关键步骤的能力，包括在疾病状态下观察到的变化。该系统的一个关键特性是使用天然丝蛋白生物材料，这使我们能够利用其生物相容性、非血栓形成特性、可编程的机械性能以及细胞因子、细胞外基质成分和内皮衍生蛋白的表面结合。这反过来为体外血液成分生产的研究提供了新机会，并为人类血小板生成病理机制的研究提供了一个优越的组织系统。
