3D Printed Bioreactors for Cell Culture

项目摘要 传统的骨损伤治疗方法有很大的局限性。而超过一百万的同种异体和 每年都会进行自体植骨手术，这会显著降低医疗并发症的发生率 -通常涉及适度的生存能力、较差的整合或免疫反应-仍然会发生。因此，灵活性 由体外培养的工程化组织提供的修复和替换 受损的骨组织。这种方法包括在支架上种植和种植细胞源，然后植入 充满细胞的结构进入损伤部位。然而，大容量工程组织的培养--以及 尤其是这些大组织中的细胞活性、扩增、增殖和分化--受到电流的限制 培养技术。为了解决这一问题，tr&d1的目标是开发一种3D打印(3DP)生物反应器作为 动态培养系统，控制细胞微环境，从而促进细胞存活，扩增， 在大型工程结构中的增殖和分化。为此，我们最近开发了一种 管状灌流系统(TPS)生物反应器，能够扩增人间充质干细胞， 这些细胞分化为成骨细胞，随后形成骨组织。基于我们的 早期的TPS生物反应器，我们将使用3D打印来制造具有可变变量的专用生物反应器 架构、受控流动环境和空间位置的细胞种群；因此，我们可以确保 足够的营养和氧气用于干细胞在这些大型结构中的扩张。 此外，3D打印控制细胞群体的空间位置将使我们能够确定相互作用 在多种细胞群之间，如间充质干细胞和内皮细胞。最后，我们将利用 在体外3D生物反应室中开发的制造可拆卸、可生物降解支架的策略 适合体内应用的工程化骨组织。这些研究的结果将提供一个 3DP生物反应器系统，可以支持大型工程组织的生长，同时还提供一套 开发其他类似设计的组织特定生物反应器系统的工具。

Project Summary Traditional treatments for bone injuries have significant limitations. While over one million allogenic and autologous bone grafting procedures are performed each year, significant incidences of medical complications - often involving modest viability, poor integration, or an immune response - still occur. Therefore, the flexibility provided by an in vitro cultured, engineered tissue provides an excellent avenue to repair and replace damaged bone tissue. This approach involves seeding and growing a cell source on a scaffold and implanting the cell-laden construct into the injury site. However, the culture of large volume engineered tissues - and particularly cell viability, expansion, proliferation, and differentiation in these large tissues - is limited by current culture techniques. To address this concern, TR&D1 aims to develop a 3D printed (3DP) bioreactor as a dynamic culture system to control cellular microenvironment and therefore promote cell viability, expansion, proliferation, and differentiation within large engineered constructs. To this end, we have recently developed a tubular perfusion system (TPS) bioreactor that enables the expansion of human mesenchymal stem cells, the differentiation of these cells into osteoblasts, and the subsequent formation of boney tissue. Based on our earlier TPS bioreactor, we will use 3D printing to fabricate specialized bioreactor chambers with variable architecture, controlled flow environments, and spatially located cell populations; thus, we can ensure adequate availability of nutrients and oxygen for the expansion of stem cells within these large constructs. Furthermore, 3D printing control of the spatial location of cell populations will allow us to determine interactions between multiple cell populations, such as mesenchymal stem cells and endothelial cells. Finally, we will utilize the strategies developed in the in vitro 3D bioreactor chambers to fabricate removable, biodegradable scaffolds of engineered bone tissues that are suitable for in vivo application. The results of these studies will deliver a 3DP bioreactor system that can support the growth of large engineered tissues, while also providing a set of tools to develop other, similarly designed, tissue specific bioreactor systems.