Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, Iran; Minimally Invasive Surgery Research Center, Human and Animal Cell Bank, Iranian Biological Resource Center, Iran University of Medical Sciences, Tehran, Iran.
Mater Sci Eng C Mater Biol Appl. 2019 Jul;100:330-340. doi: 10.1016/j.msec.2019.02.090. Epub 2019 Feb 26.
Cancer is the second leading cause of human death. Therefore, comprehensive research and the appropriate tools are needed in this field. Animal models and cell culture studies are the most important preclinical tools in cancer research. In 2D cell culture models, cells are forced to grow in a 2D environment, which differs from their natural physiology. Recently, 3D cell culture models were developed to fill the gap between 2D cell culture and animal models.
Human amniotic membranes were obtained, decellularized, characterized and used as a natural 3D scaffold to investigate cancer cell behavior in 2D compared to 3D conditions. Time-lapse imaging of cells was used, and cell proliferation, velocity and migration were evaluated. Cisplatin was applied in 2D and 3D conditions, followed by evaluation of viability, apoptosis and cancer stem cell proteins by flow cytometry and western blot analysis.
The results showed that in the decellularized amnion membrane (DAM) scaffold most cells did not spread and remain rounded and then penetrated into the scaffold with no cytotoxicity. Significant differences in migration, velocity, morphology and proliferation of cancer cells were observed between the 3D DAM scaffold and 2D model. Furthermore, the cells in the 3D DAM scaffold showed much more resistance to apoptosis and higher CSC content.
In conclusion, considering the effect of the 3D DAM scaffold in cell behavior, apoptosis resistance and CSC content as well as the short processing time for decellularizing the AM, it appears that the 3D DAM scaffold offers an appropriate tool for in vitro cancer research.
癌症是人类死亡的第二大主要原因。因此，该领域需要全面的研究和适当的工具。动物模型和细胞培养研究是癌症研究中最重要的临床前工具。在 2D 细胞培养模型中，细胞被迫在 2D 环境中生长，这与它们的自然生理状态不同。最近，开发了 3D 细胞培养模型来填补 2D 细胞培养和动物模型之间的空白。
获得人羊膜，去细胞化，进行特征分析，并用作天然 3D 支架，以研究 2D 条件下与 3D 条件下的癌细胞行为。使用细胞延时成像，评估细胞增殖、速度和迁移。在 2D 和 3D 条件下应用顺铂，然后通过流式细胞术和 Western blot 分析评估细胞活力、细胞凋亡和癌症干细胞蛋白。
结果表明，在去细胞化羊膜（DAM）支架中，大多数细胞没有扩散，保持圆形，然后穿透支架，没有细胞毒性。在 3D DAM 支架和 2D 模型之间观察到癌细胞迁移、速度、形态和增殖的显著差异。此外，3D DAM 支架中的细胞对细胞凋亡的抵抗力更强，CSC 含量更高。
总之，考虑到 3D DAM 支架对细胞行为、细胞凋亡抵抗和 CSC 含量的影响，以及去细胞化 AM 的短处理时间，3D DAM 支架似乎为体外癌症研究提供了一种合适的工具。
