Department of Basic Pharmaceutical Sciences, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey.
Bioengineering Division, Institute for Graduate Studies in Science and Engineering, Hacettepe University, Ankara, Turkey.
Biotechnol Bioeng. 2020 Aug;117(8):2527-2539. doi: 10.1002/bit.27381. Epub 2020 May 29.
Three-dimensional (3D) cancer tumor models are becoming vital approaches for high-throughput drug screening, drug targeting, development of novel theranostic systems, and personalized medicine. Yet, it is becoming more evident that the tumor progression and metastasis is fueled by a subpopulation of stem-like cells within the tumor that are also called cancer stem cells (CSCs). This study aimed to develop a tumoroid model using CSCs. For this purpose CD133 cells were isolated from SaOS-2 osteosarcoma cell line with magnetic-activated cell sorting. To evaluate tumoroid formation ability, the cells were incubated in different cell numbers in agar gels produced by 3D Petri Dish® method. Subsequently, CD133 cells and CD133 cells were co-cultured to investigate CD133 cell localization in tumoroids. The characterization of tumoroids was performed using Live&Dead staining, immunohistochemistry, and quantitative polymerase chain reaction analysis. The results showed that, CD133 , CD133 and SaOS-2 cells were all able to form 3D tumoroids regardless of the initial cell number, but, while 72 hr were needed for CD133 cells to self-assemble, 24 hr were enough for CD133 and SaOS-2 cells. CD133 cells were located within tumoroids randomly with high cell viability. Finally, when compared to two-dimensional (2D) cultures, there were 5.88, 4.14, 6.95, and 1.68-fold higher messenger RNA expressions for Sox2, OCT3/4, Nanog, and Nestin, respectively, in CD133 cells that were cultured within 3D tumoroids, showing longer maintenance of stem cell phenotype in 3D, that can allow more relevant screening and targeting efficiency in pharmaceutical testing. It was concluded that CSC-based tumoroids are propitious as 3D tumor models to fill the gap between conventional 2D in vitro culture and in vivo animal experiments for cancer research.
三维（3D）癌症肿瘤模型正成为高通量药物筛选、药物靶向、新型治疗诊断系统开发和个性化医疗的重要方法。然而，越来越明显的是，肿瘤的进展和转移是由肿瘤内的一小部分干细胞样细胞（也称为癌症干细胞（CSC））驱动的。本研究旨在使用 CSC 开发肿瘤模型。为此，使用磁性激活细胞分选从 SaOS-2 骨肉瘤细胞系中分离 CD133 细胞。为了评估肿瘤形成能力，将细胞在 3D Petri 盘®方法产生的琼脂凝胶中以不同的细胞数孵育。随后，将 CD133 细胞和 CD133 细胞共培养，以研究 CD133 细胞在肿瘤中的定位。通过 Live&Dead 染色、免疫组织化学和定量聚合酶链反应分析对肿瘤进行表征。结果表明，无论初始细胞数如何，CD133+、CD133 和 SaOS-2 细胞都能够形成 3D 肿瘤，但 CD133 细胞需要 72 小时自我组装，而 CD133 和 SaOS-2 细胞则需要 24 小时。CD133 细胞随机位于肿瘤内，细胞活力高。最后，与二维（2D）培养相比，在 3D 肿瘤中培养的 CD133 细胞的 Sox2、OCT3/4、Nanog 和 Nestin 的信使 RNA 表达分别高出 5.88、4.14、6.95 和 1.68 倍，表明在 3D 中干细胞表型的维持时间更长，这可以在药物测试中提高更相关的筛选和靶向效率。研究得出结论，基于 CSC 的肿瘤是有利的 3D 肿瘤模型，可以填补传统 2D 体外培养和体内动物实验之间的空白，用于癌症研究。
