Biopharmaceutical Research Center, Ochang Institute of Biological and Environmental Science, Korea Basic Science Institute (KBSI), Cheongju 28119, Republic of Korea.
Korea University of Science and Technology (UST), Daejeon 34113, Republic of Korea.
ACS Appl Bio Mater. 2024 Nov 18;7(11):7194-7206. doi: 10.1021/acsabm.4c00800. Epub 2024 Oct 11.
Cell-based immunotherapies have emerged as promising cancer treatment modalities, demonstrating remarkable clinical efficacy. As interest in applying immune cell-based therapies to solid tumors has gained momentum, experimental models that enable long-term monitoring and mimic clinical administration are increasingly necessary. This study explores the potential of scaffold-based cell culture technologies, specifically three-dimensional (3D) extracellular matrix (ECM)-like frameworks, as promising solutions. These frameworks facilitate unhindered immune cell growth and enable continuous cancer cell culture. The three-dimensional (3D) cell culture model was developed using tailored scaffolds for natural killer (NK) cell culture. Within this framework, A549 lung cancer cells were cocultured with NK cells, allowing real-time monitoring for up to 28 days. The expression of critical markers associated with anticancer drug resistance and epithelial-mesenchymal transition (EMT) was evaluated in cancer cells within this 3D culture context. Compared to conventional 2D monolayer cultures, this 3D scaffold-based culture revealed that solid tumor cells, specifically A549 cells, exhibited heightened resistance to anticancer drugs. Additionally, the 3D culture environment upregulated the expression of EMT markers namely vimentin, N-cadherin, and fibronectin, while NK and zEGFR-CAR-NK cells displayed anticancer effects. In the two-dimensional (2D) coculture, only zEGFR-CAR-NK cells exhibited such effects in the 3D coculture system, highlighting an intriguing inconsistency with the 2D culture model, further confirmed by experiments. This 3D cell culture model reliably predicts outcomes in NK immunotherapy experiments. Thus, it represents a valuable tool for investigating drug resistance mechanisms and assessing the efficacy of immune cell-based therapies. By bridging the gap between and investigations, this model effectively translates potential treatments into animal models and facilitates rigorous preclinical evaluations.
基于细胞的免疫疗法已成为有前途的癌症治疗方式，显示出显著的临床疗效。随着将免疫细胞为基础的疗法应用于实体瘤的兴趣不断增加，越来越需要能够进行长期监测并模拟临床给药的实验模型。本研究探讨了支架为基础的细胞培养技术的潜力，特别是三维（3D）细胞外基质（ECM）样框架，作为有前途的解决方案。这些框架促进免疫细胞的无阻碍生长，并实现持续的癌细胞培养。使用专门设计的支架开发了用于自然杀伤（NK）细胞培养的 3D 细胞培养模型。在这个框架内，将 A549 肺癌细胞与 NK 细胞共培养，可实时监测长达 28 天。在这个 3D 培养环境中评估了与抗癌药物耐药性和上皮-间充质转化（EMT）相关的关键标志物在癌细胞中的表达。与传统的 2D 单层培养相比，这种 3D 支架培养揭示了实体瘤细胞，特别是 A549 细胞，对抗癌药物表现出更高的耐药性。此外，3D 培养环境上调了 EMT 标志物的表达，即波形蛋白、N-钙黏蛋白和纤连蛋白，而 NK 和 zEGFR-CAR-NK 细胞则表现出抗癌作用。在二维（2D）共培养中，只有 zEGFR-CAR-NK 细胞在 3D 共培养系统中表现出这种作用，这与 2D 培养模型不一致，进一步通过实验得到证实。这种 3D 细胞培养模型可靠地预测了 NK 免疫治疗实验的结果。因此，它是研究药物耐药机制和评估免疫细胞为基础的治疗方法疗效的有价值工具。通过弥合基础和临床研究之间的差距，该模型有效地将潜在的治疗方法转化为动物模型，并促进严格的临床前评估。
