Institute of Cell Biology and Biophysics, Leibniz University Hannover, Hannover, Germany.
Institute of Cell and Tissue Culture Technologies, Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria.
Sci Rep. 2024 Apr 30;14(1):9904. doi: 10.1038/s41598-024-60822-z.
Animal models lack physiologic relevance to the human system which results in low clinical translation of results derived from animal testing. Besides spheroids or organoids, hydrogel-based 3D in vitro models are used to mimic the in vivo situation increasing the relevance while reducing animal testing. However, to establish hydrogel-based 3D models in applications such as drug development or personalized medicine, high-throughput culture systems are required. Furthermore, the integration of oxygen-reduced (hypoxic) conditions has become increasingly important to establish more physiologic culture models. Therefore, we developed a platform technology for the high-throughput generation of miniaturized hydrogels for 3D cell culture. The Oli-Up system is based on the shape of a well-plate and allows for the parallel culture of 48 hydrogel samples, each with a volume of 15 µl. As a proof-of-concept, we established a 3D culture of gelatin-methacryloyl (GelMA)-encapsulated mesenchymal stem/stromal cells (MSCs). We used a hypoxia reporter cell line to establish a defined oxygen-reduced environment to precisely trigger cellular responses characteristic of hypoxia in MSCs. In detail, the expression of hypoxia response element (HRE) increased dependent on the oxygen concentration and cell density. Furthermore, MSCs displayed an altered glucose metabolism and increased VEGF secretion upon oxygen-reduction. In conclusion, the Oli-Up system is a platform technology for the high-throughput culture of hydrogel-based 3D models in a defined oxygen environment. As it is amenable for automation, it holds the potential for high-throughput screening applications such as drug development and testing in more physiologic 3D in vitro tissue models.
动物模型缺乏与人类系统的生理相关性，导致从动物试验中得出的结果在临床上的转化较低。除了球体或类器官外，基于水凝胶的 3D 体外模型也被用于模拟体内情况，从而提高相关性，同时减少动物试验。然而，为了在药物开发或个性化医疗等应用中建立基于水凝胶的 3D 模型，需要高通量培养系统。此外，缺氧（低氧）条件的整合对于建立更生理的培养模型变得越来越重要。因此，我们开发了一种用于高通量生成用于 3D 细胞培养的微型水凝胶的平台技术。Oli-Up 系统基于微孔板的形状，允许并行培养 48 个水凝胶样本，每个样本的体积为 15μl。作为概念验证，我们建立了明胶甲基丙烯酰（GelMA）包封间充质干细胞/基质细胞（MSCs）的 3D 培养。我们使用缺氧报告细胞系来建立定义的低氧环境，以精确触发 MSCs 中缺氧的特征性细胞反应。具体而言，缺氧反应元件（HRE）的表达随氧浓度和细胞密度的增加而增加。此外，MSCs 在氧还原时表现出改变的葡萄糖代谢和增加的 VEGF 分泌。总之，Oli-Up 系统是一种用于在定义的氧环境中高通量培养基于水凝胶的 3D 模型的平台技术。由于它适合自动化，因此具有在更生理的 3D 体外组织模型中进行高通量筛选应用的潜力，例如药物开发和测试。
