Wang Yaqi, Tian Feng, Duan Yujie, Li Zhuogang, Chen Qiaotong, Chen Jiaying, Miao Xiaomin, Xu Yanhong, Xu Chulan, Tang Yadong
School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, China.
School of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou 510006, China.
Tissue Cell. 2022 Jun;76:101796. doi: 10.1016/j.tice.2022.101796. Epub 2022 Apr 6.
The development of three-dimensional (3D) in vitro model to recapitulate the in vivo tumor tissue is essential for studying tumor biology, discovering anti-cancer drugs, and evaluating anti-cancer drug efficacy. However, most of the previous models lack the involvement of vascular barrier. Here, we proposed an in vitro 3D cocultured tumor-vascular barrier model by the combination of alginate hydrogels beads and Transwell system. PC-3 cells and NIH/3T3 cells were encapsulated in alginate hydrogel beads, which were cultured in the bottom chamber of Transwell, while human umbilical vein endothelial cells (HUVECs) were cultured on the porous membrane in the upper chamber to form vascular barrier. The effect of the concentration of alginate sodium on the morphology, diameter and swelling ratio of the beads was studied. The alginate sodium content and cell seeding density were further optimized according to cell proliferation ability. The formation of endothelial barrier was verified by immunostaining with tight junction protein VE-cadherin and transendothelial electrical resistance (TEER) monitoring. Finally, the drug response of 3D cocultured tumor-vascular barrier model to curcumin was evaluated. Compared with two-dimensional (2D) coculture model and 3D coculture spheroid model, 3D tumor-vascular barrier model showed the highest activity of cancer cells and the strongest drug resistance. The developed 3D cocultured tumor-vascular barrier model possesses great potential to be applied for in vitro evaluation of anti-tumor drugs.
构建能够模拟体内肿瘤组织的三维（3D）体外模型对于研究肿瘤生物学、发现抗癌药物以及评估抗癌药物疗效至关重要。然而，大多数先前的模型缺乏血管屏障的参与。在此，我们通过藻酸盐水凝胶珠与Transwell系统相结合，提出了一种体外3D共培养肿瘤-血管屏障模型。将PC-3细胞和NIH/3T3细胞封装在藻酸盐水凝胶珠中，在Transwell的底部腔室中培养，而人脐静脉内皮细胞（HUVECs）在上部腔室的多孔膜上培养以形成血管屏障。研究了海藻酸钠浓度对珠子形态、直径和溶胀率的影响。根据细胞增殖能力进一步优化了海藻酸钠含量和细胞接种密度。通过紧密连接蛋白VE-钙黏蛋白的免疫染色和跨内皮电阻（TEER）监测验证了内皮屏障的形成。最后，评估了3D共培养肿瘤-血管屏障模型对姜黄素的药物反应。与二维（2D）共培养模型和3D共培养球体模型相比，3D肿瘤-血管屏障模型显示出最高的癌细胞活性和最强的耐药性。所构建的3D共培养肿瘤-血管屏障模型在体外抗肿瘤药物评价方面具有巨大的应用潜力。
