Key Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education, Institute for Clean Energy and Advanced Materials, School of Materials and Energy, Southwest University, Chongqing 400715, PR China.
College of Chemistry and Chemical Engineering, Hainan Normal University, Haikou 571158, PR China.
Biomater Adv. 2023 Oct;153:213507. doi: 10.1016/j.bioadv.2023.213507. Epub 2023 Jun 8.
Scaffold-based culture is one of the effective methods to resemble three-dimensional (3D) cells model in vitro. An agar@lens paper hybrid scaffold was prepared by one-pot dip-coating. The lens paper's cellulose fiber networks are the scaffold's backbone. The agar gel seized the gaps between the fibrous structures that can improve the paper scaffold's optical transparency and prevent cells from spreading on the scaffold. The SEM and light microscope images showed that the agar gel on the bottom of the paper and the cellulose fiber of the paper formed micro-well structures. Without staining, the cells growing on the agar@paper scaffold can be directly observed under a light microscope. Cells aggregated between the cellulose fibers and formed spheroids within 24 h. The cell spheroids can be non-enzymatically retrieved from the agar@paper scaffold because of the cell-repelling property of agar. The agar@paper scaffold was applied for co-culturing tumor cells (MDA-MB-231, DU 145) and natural killer cells (NKs, NK-92). Using the agar@paper scaffolds, the tumor-infiltrating NKs can be separated from floating NKs that did not attack the tumor spheroids. The effect of NKs infiltrating on tumor spheroids size was characterized. The results showed that NKs attacking the spheroids grown on agar@paper scaffold can be readily tracked because of the improved optical transparency. Higher NKs: tumor cells ratio resulted in a high percentage of tumor infiltrating NKs. The separated NKs can be further tested to reveal their biological characteristics. Both agar and lens paper is accessible for most biological labs, highlighting the potential of agar@paper scaffold in 3D culture.
支架培养是体外模拟三维（3D）细胞模型的有效方法之一。通过一锅浸渍法制备了琼脂@透镜纸杂化支架。透镜纸的纤维素纤维网络是支架的骨架。琼脂凝胶占据纤维结构之间的间隙，可以提高纸支架的光学透明度并防止细胞在支架上扩散。SEM 和光学显微镜图像显示，纸底部的琼脂凝胶和纸的纤维素纤维形成微井结构。未经染色，在光镜下可直接观察到在琼脂@纸上支架上生长的细胞。细胞在 24 小时内聚集在纤维素纤维之间并形成球体。由于琼脂的细胞排斥特性，细胞球体可以从琼脂@纸上支架中非酶回收。琼脂@纸支架用于共培养肿瘤细胞（MDA-MB-231、DU 145）和自然杀伤细胞（NKs、NK-92）。使用琼脂@纸支架，可以将浸润肿瘤的 NK 从未攻击肿瘤球体的漂浮 NK 中分离出来。还对 NK 浸润对肿瘤球体大小的影响进行了表征。结果表明，由于光学透明度的提高，可轻松跟踪攻击琼脂@纸上支架上球体的 NK。较高的 NK：肿瘤细胞比例导致较高比例的肿瘤浸润 NK。分离的 NK 可以进一步测试以揭示其生物学特性。琼脂和透镜纸都可供大多数生物实验室使用，突出了琼脂@纸支架在 3D 培养中的潜力。
