Janvier Adam J, Canty-Laird Elizabeth, Henstock James R
Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, UK.
J Tissue Eng. 2020 Sep 1;11:2041731420942462. doi: 10.1177/2041731420942462. eCollection 2020 Jan-Dec.
A range of bioreactors use linear actuators to apply tensile forces , but differences in their culture environments can limit a direct comparison between studies. The widespread availability of 3D printing now provides an opportunity to develop a 'universal' bioreactor chamber that, with minimal exterior editing can be coupled to a wide range of commonly used linear actuator platforms, for example, the EBERS-TC3 and CellScale MCT6, resulting in a greater comparability between results and consistent testing of potential therapeutics. We designed a bioreactor chamber with six independent wells that was 3D printed in polylactic acid using an Ultimaker 2+ and waterproofed using a commercially available coating (XTC-3D), an oxirane resin. The cell culture wells were further coated with Sylgard-184 polydimethylsiloxane (PDMS) to produce a low-adhesion well surface. With appropriate coating and washing steps, all materials were shown to be non-cytotoxic by lactate dehydrogenase assay, and the bioreactor was waterproof, sterilisable and reusable. Tissue-engineered tendons were generated from human mesenchymal stem cells in a fibrin hydrogel and responded to 5% cyclic strain (0.5 Hz, 5 h/day, 21 days) in the bioreactor by increased production of collagen-Iα1 and decreased production of collagen-IIIα1. Calcification of the extracellular matrix was observed in unstretched tendon controls indicating abnormal differentiation, while tendons cultured under cyclic strain did not calcify and exhibited a tenogenic phenotype. The ease of manufacturing this bioreactor chamber enables researchers to quickly and cheaply reproduce this culture environment for use with many existing bioreactor actuator platforms by downloading the editable CAD files from a public database and following the manufacturing steps we describe.
一系列生物反应器使用线性致动器来施加拉力，但它们培养环境的差异可能会限制不同研究之间的直接比较。3D打印技术的广泛应用现在提供了一个机会，来开发一种“通用”的生物反应器腔室，通过最少的外部编辑，它可以与各种常用的线性致动器平台（例如EBERS-TC3和CellScale MCT6）耦合，从而使结果之间具有更大的可比性，并能对潜在疗法进行一致的测试。我们设计了一个带有六个独立孔的生物反应器腔室，使用Ultimaker 2+ 3D打印机以聚乳酸打印而成，并使用一种市售涂层（XTC-3D，一种环氧乙烷树脂）进行防水处理。细胞培养孔进一步用Sylgard-184聚二甲基硅氧烷（PDMS）涂层，以产生低粘附性的孔表面。通过适当的涂层和清洗步骤，乳酸脱氢酶测定表明所有材料均无细胞毒性，并且该生物反应器具有防水、可消毒和可重复使用的特性。在纤维蛋白水凝胶中利用人间充质干细胞生成了组织工程化肌腱，并且在生物反应器中，这些肌腱对5%的循环应变（0.5 Hz，每天5小时，共21天）产生反应，表现为I型胶原α1的产量增加和III型胶原α1的产量减少。在未拉伸的肌腱对照中观察到细胞外基质钙化，表明分化异常，而在循环应变下培养的肌腱未发生钙化，并表现出成腱细胞表型。通过从公共数据库下载可编辑的CAD文件并按照我们描述的制造步骤，制造这种生物反应器腔室的简便性使研究人员能够快速且廉价地重现这种培养环境，以便与许多现有的生物反应器致动器平台配合使用。
