Berg Johanna, Heinze Julian, Niemeyer Daniela, Hellgren Josefin, Jaiswal Himjyot, Löwa Anna, Hocke Andreas, Namro Itedale, Drosten Christian, Kurreck Jens, Tolksdorf Beatrice
Department of Applied Biochemistry, Institute of Biotechnology, Technische Universität Berlin, 10623 Berlin, Germany.
German Center for Infection Research (DZIF), Charitéplatz 1, 10117 Berlin, Germany.
Int J Mol Sci. 2025 Jun 10;26(12):5543. doi: 10.3390/ijms26125543.
Viral lung infections are a never-ending threat to public health due to the emergence of new variants and their seasonal nature. While vaccines offer some protection, the need for effective antiviral drugs remains high. The existing research methods using 2D cell culture and animal models have their limitations. Human cell-based tissue engineering approaches hold great promise for bridging this gap. Here, we describe a microextrusion bioprinting approach to generate three-dimensional (3D) lung models composed of four cell types: endothelial cells, primary fibroblasts, macrophage cells, and epithelial cells. A549 and Calu-3 cells were selected as epithelial cells to simulate the cells of the lower and upper respiratory tract, respectively. Cells were bioprinted in a hydrogel consisting of alginate, gelatin, hyaluronic acid, collagen, and laminin-521. The models were cultured under air-liquid interface (ALI) conditions to further enhance their physiological relevance as lung cells. Their viability, metabolic activity, and expression of specific cell markers were analyzed during long-term culture for 21 days. The constructs were successfully infected with both a seasonal influenza A virus (IAV) and the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) omicron variant, demonstrating their potential for studying diverse viral infections.
由于新变种的出现及其季节性特征，病毒性肺部感染对公众健康构成了持续不断的威胁。虽然疫苗提供了一定程度的保护，但对有效抗病毒药物的需求仍然很高。现有的使用二维细胞培养和动物模型的研究方法存在局限性。基于人类细胞的组织工程方法在弥合这一差距方面具有巨大潜力。在此，我们描述了一种微挤压生物打印方法，用于生成由四种细胞类型组成的三维（3D）肺模型：内皮细胞、原代成纤维细胞、巨噬细胞和上皮细胞。分别选择A549和Calu-3细胞作为上皮细胞，以模拟下呼吸道和上呼吸道的细胞。细胞在由藻酸盐、明胶、透明质酸、胶原蛋白和层粘连蛋白-521组成的水凝胶中进行生物打印。这些模型在气液界面（ALI）条件下培养，以进一步增强其作为肺细胞的生理相关性。在长达21天的长期培养过程中，分析了它们的活力、代谢活性和特定细胞标志物的表达。构建体成功感染了季节性甲型流感病毒（IAV）和严重急性呼吸综合征冠状病毒2（SARS-CoV-2）奥密克戎变种，证明了它们在研究多种病毒感染方面的潜力。
