Application of additive manufacturing technologies for the realization of clinically relevant cocultures of mammalian cells and microalgae

应用增材制造技术实现临床相关的哺乳动物细胞和微藻共培养

• 批准号: 417020100
• 负责人: Professor Dr.-Ing. Felix Krujatz
• 金额: --
• 依托单位: Zentrum für Translationale Knochen-, Gelenk- und Weichgewebeforschung
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/417020100?language=en
• 关键词: Application; additive; manufacturing; technologies; realization

The regeneration of lost tissue functions by transplantation of cells, often in combination with biomaterials as artificial matrix, is a promising approach for the treatment of various diseases and tissue defects. However, the success of cell-based therapies strongly depends on the supply of the cells, especially with oxygen, immediately after implantation. For most tissue engineering approaches, strategies for a fast revascularization are subject of research. On the other side, there are treatment approaches which intend to circumvent contact of the implanted cells with immunological components in blood in order to avoid rejection and therefore need alternative strategies for efficient oxygen supply. One example of high clinical relevance is the transplantation of allogenic (potentially xenogenic) pancreatic islets of Langerhans in case of patients suffering from diabetes type 1. First short-term experiments demonstrated the great potential of the utilization of photosynthetically active microalgae for coverage of the high oxygen demand of pancreatic islets.In the proposed project, cocultures of photosynthetically active microalgae and mammalian cells will be realized by means of bioprinting. Both cell types will be encapsulated (separated from each other) in hydrogels; by applying the additive manufacturing technology of 3D plotting for shaping, the advantages of a spatially defined arrangement of the different cell types in a structured hydrogel matrix with defined geometry will be used. Beyond that, a bioreactor system with light coupling and integrated oxygen sensors will be developed that allows a demand-oriented control of the photosynthesis activity depending on the actual oxygen concentration. With the coculture/bioreactor system the fundamentals will be developed for a systematic investigation of the interaction between the different cell types, the influence of the oxygen concentration on the cells as well as the oxygen transport between the different compartments. Moreover, an efficient and long-lasting supply of the mammalian cells with photosynthetically generated oxygen should be achieved as prerequisite for the translation of the coculture/bioreactor system towards clinical application for islet transplantation. For the establishment of the coculture, a suitable microalgae strain should be identified which is able to growth at 37°C and is characterized by a high rate of photosynthesis. Coculture conditions will be worked out which meet the demands of both cell types. The influence of organic carbon sources on the photoautotrophic microalgae as well as the influence of light on the heterotrophic mammalian cells will be investigated in detail. Finally, it is expected that results of the project concerning the bioprinting with microalgae will provide valuable insights for biotechnological applications.

通过细胞移植来再生失去的组织功能，通常与生物材料作为人工基质结合，是治疗各种疾病和组织缺陷的一种很有前途的方法。然而，细胞疗法的成功很大程度上取决于植入后细胞的供应，尤其是氧气的供应。对于大多数组织工程方法，快速血运重建的策略是研究的主题。另一方面，有一些治疗方法旨在避免植入细胞与血液中的免疫成分接触，以避免排斥反应，因此需要替代有效供氧的策略。一个具有高度临床相关性的例子是同种异体（可能是异种）朗格汉斯胰岛移植，用于治疗1型糖尿病患者。最初的短期实验证明了利用光合活性微藻覆盖胰岛高需氧量的巨大潜力。在本项目中，将通过生物打印技术实现光合活性微藻与哺乳动物细胞的共培养。两种类型的细胞都将被包裹在水凝胶中（彼此分离）；通过应用3D绘图的增材制造技术进行成形，将利用具有定义几何形状的结构化水凝胶矩阵中不同细胞类型的空间定义排列的优势。除此之外，将开发一种具有光耦合和集成氧传感器的生物反应器系统，该系统可以根据实际氧浓度以需求为导向控制光合作用活动。随着共培养/生物反应器系统的基础将被发展为不同细胞类型之间的相互作用，氧浓度对细胞的影响以及不同隔间之间的氧运输的系统研究。此外，有效和持久地为哺乳动物细胞提供光合作用产生的氧气是将共培养/生物反应器系统转化为胰岛移植临床应用的先决条件。为了建立共培养，应该确定一种适合的微藻菌株，该菌株能够在37℃下生长，并且具有高光合速率的特点。将制定出满足两种细胞类型要求的共培养条件。本文将详细研究有机碳源对光自养微藻的影响以及光对异养哺乳动物细胞的影响。最后，期望微藻生物打印项目的成果将为生物技术应用提供有价值的见解。