Jun Brian H, Torrez Jacob E, Ross David J, Patterson Brian M, Ishak Mohammad O, Rodriguez Arasely M, Harris Jennifer F, Davis-Anderson Katie L
Biochemistry and Biotechnology Group, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Engineered Materials Group, Material Science and Technology Division, Los Alamos National Laboratory, Los Alamos, NM, USA.
Sci Rep. 2025 Mar 24;15(1):10134. doi: 10.1038/s41598-025-94093-z.
We introduce a novel fabrication method for developing a 3D-printed perfusion bioreactor (3D-PBR) to facilitate the in situ growth and differentiation of human bone marrow (BM)-derived mesenchymal stem cells (MSCs) while enabling coculture with vascular cells. To recapitulate human physiology, in vitro platforms must incorporate several key features of their native target organ. This often entails a supportive 3D architecture for growing and differentiating multiple human cell types in situ under perfusion. Other essential characteristics include reproducibility, ease of customization, and biocompatibility. Our 3D-PBR combines these features and was fabricated using a biocompatible resin-based polymer, which was 3D-printed, followed by the addition of a permeable membrane to create a coculture microenvironment. MSCs were encapsulated in a collagen-fibrin gel alongside human endothelium within the 3D-PBR. The physical cues that our 3D-PBR provided facilitated the differentiation of MSCs into specific lineages, such as adipocytes and osteoblasts. Immunohistochemistry images demonstrated that cells grown in the 3D-PBR exhibited more physiologically relevant BM perivascular niche markers compared to static culture models. Our method utilizes emerging 3D printing techniques and alternative materials, departing from traditional PDMS-based soft lithography. These advancements in fabrication further enhance in vitro platforms for diverse cell culture models and vascular permeability assays.
我们介绍了一种用于开发3D打印灌注生物反应器（3D-PBR）的新颖制造方法，以促进人骨髓（BM）来源的间充质干细胞（MSC）的原位生长和分化，同时实现与血管细胞的共培养。为了重现人体生理过程，体外平台必须具备其天然靶器官的几个关键特征。这通常需要一个支持性的3D架构，以便在灌注条件下原位培养和分化多种人类细胞类型。其他基本特征包括可重复性、易于定制和生物相容性。我们的3D-PBR结合了这些特征，并使用基于生物相容性树脂的聚合物制造，该聚合物经过3D打印，然后添加可渗透膜以创建共培养微环境。在3D-PBR内，MSC与人类内皮细胞一起被封装在胶原蛋白-纤维蛋白凝胶中。我们的3D-PBR提供的物理线索促进了MSC向特定谱系（如脂肪细胞和成骨细胞）的分化。免疫组织化学图像显示，与静态培养模型相比，在3D-PBR中生长的细胞表现出更多与生理相关的BM血管周围生态位标记物。我们的方法采用了新兴的3D打印技术和替代材料，有别于传统的基于聚二甲基硅氧烷（PDMS）的软光刻技术。这些制造方面的进步进一步增强了用于各种细胞培养模型和血管通透性测定的体外平台。
